Phenyl amidine alkanoic acids and lactones useful as platelet aggregation inhibitors

ABSTRACT

This invention relates to phenyl amidine alkanoic acids and lactones having the following formula ##STR1## or a pharmaceutically acceptable salt which are useful in the inhibition of platelet aggregation. This invention also relates to pharmaceutical compositions of such phenyl amidine derivatives.

This application is a 371 of PCT/US93/05861, filed Jun. 23, 1993 nowWO94/00424 issued Jan. 6, 1994 which is a CIP of Ser. No. 07/904,237 nowabandoned.

FIELD OF THE INVENTION

This invention is in the field of mammalian therapeutics and relates tocompounds for the treatment of mammalian disorders such ascardiovascular disorders. Of particular interest is a class of phenylamidines derivatives useful as inhibitors of platelet aggregation.

BACKGROUND OF THE INVENTION

Fibrinogen is a glycoprotein present as a normal component of bloodplasma. It participates in platelet aggregation and fibrin formation inthe blood clotting mechanism.

Platelets are cellular elements found in whole blood which alsoparticipate in blood coagulation. Fibrinogen binding to platelets isimportant to normal platelet function in the blood coagulationmechanism. When a blood vessel receives an injury, the platelets bindingto fibrinogen will initiate aggregation and form a thrombus. Interactionof fibrinogen with platelets occurs through a membrane glycoproteincomplex, known as gpIIb/IIIa; this is an important feature of theplatelet function. Inhibitors of this interaction are useful inmodulating platelet thrombus formation.

It is also known that another large glycoprotein named fibronectin,which is a major extracellular matrix protein, interacts with fibrinogenand fibrin, and with other structural molecules such as actin, collagenand proteoglycans. Various relatively large polypeptide fragments in thecell-binding domain of fibronectin have been found to havecell-attachment activity. See U.S. Pat. Nos. 4,517,686; 4,589,881; and4,661,111. Certain relatively short peptide fragments from the samemolecule were found to promote cell attachment to a substrate whenimmobilized on the substrate or to inhibit attachment when in asolubilized or suspended form. See U.S. Pat. Nos. 4,578,079 and4,614,517.

In U.S. Pat. No. 4,683,291, inhibition of platelet function is disclosedwith synthetic peptides designed to be high affinity antagonists offibrinogen binding to platelets. U.S. Pat. No. 4,857,508 disclosestetrapeptides having utility as inhibitors of platelet aggregation.

Other synthetic peptides and their use as inhibitors of fibrinogenbinding to platelets are disclosed by Koczewiak et al., Biochem. 23,1767-1774 (1984); Plow et al., Proc. Natl. Acad. Sci. 82, 8057-8061(1985); Ruggeri et al., Ibid. 83, 5708-5712 (1986); Ginsberg et al., J.Biol. Chem. 260 (7), 3931-3936 (1985); Hayerstick et al., Blood 66 (4),946-952 (1985); and Ruoslahti and Pierschbacher, Science 238, 491-497(1987). Still other such inhibitory peptides are disclosed in EP PatentApplications 275,748 and 298,820.

U.S. Pat. No. 4,879,313 discloses compounds useful as inhibitors ofplatelet aggregation having the formula: ##STR2## wherein x=6 to 10,

y=0 to 4,

Z=H, COOH, CONH₂ OR C₁₋₆ alkyl,

Ar=phenyl, biphenyl or naphthyl, each substituted with 1 to 3 methoxygroups, or an unsubstituted phenyl, biphenyl, naphthyl, pyridyl orthienyl group, and

Asp=aspartic acid residue.

U.S. Pat. No. 4,977,168 discloses compounds having the followingstructural formula ##STR3## wherein R₁ represents hydrogen, a loweralkyl group, a lower hydroxyalkyl group, a benzyl group, a phenyl groupor a 4-hydroxyphenyl group;

R₂ represents a lower alkyl, lower alkenyl, lower alkynyl or benzylgroup, or a lower alkoxycarbonylalkyl, lower carboxyalkyl, or lowerhydroxyalkyl group;

R₃ and R₄ identical or different, each represents a lower alkyl or lowerhydroxyalkyl radical, lower alkenyl or lower alkynyl radical or formtogether with the nitrogen to which they are attached, a saturatedheterocycle such as morpholino, thiomorpholino, pyrrolidino notsubstituted or substituted by an alkoxycarbonyl or carboxy group,piperazino, 4-(lower alkyl)piperazino, 4-(lower hydroxyalkyl)piperazino,or piperidino not substituted or substituted by one of the followinggroups: lower alkyl, benzyl, hydroxy, lower hydroxyalkyl, amino, loweraminoalkyl, hydroxyamino, alkoxycarbonyl or carboxy.

Ar represents a phenyl, alpha-naphthyl or beta-naphthyl group, possiblysubstituted, or a heteroaryl group chosen from the radicals pyridyl,quinolinyl, or isoquinolinyl, possibly substituted, as well as theirisomers and their mixtures and their salts with pharmaceuticallyacceptable mineral or organic acids which are useful as antithromboticagents.

U.S. Pat. No. 4,791,102 discloses compounds having the followingstructural formula ##STR4## wherein R₁ represents a lower alkyl, lowerhydroxyalkyl, or benzyl group, a phenyl or a 4-hydroxyphenyl group.

R₂ and R₃ identical or different, each represents a lower alkyl orhydroxyalkyl, lower alkenyl or lower alkynyl radical, or they formtogether with the nitrogen to which they are attached, a saturatedheterocycle such as morpholino, thiomorpholino, pyrrolidinounsubstituted or substituted by an alkoxycarbonyl or carboxyl group,piperazino, 4-(lower alkyl)-piperazino or piperidino unsubstituted orsubstituted by a lower alkyl, benzyl, hydroxy, lower hydroxyalkyl,amino, lower aminoalkyl, alkoxycarbonyl or carboxyl group.

Ar represents a phenyl, a possibly substituted alpha-naphthyl orbeta-naphthyl group, or else a heteroaryl group chosen from pyridyl,quinolinyl, isoquinolinyl, possibly substituted which are useful asselective inhibiting agents of thrombin and antithrombotics.

European Patent Application 372,486 discloses N-acyl beta amino acidderivatives of the formula: ##STR5## and their salts. Said compounds areuseful for inhibiting platelet aggregation in the treatment ofthrombosis, stroke, myocardial infarction, inflammation andarteriosclerosis, and for inhibiting metastasis.

European Patent Application 381 033 A1 discloses amidino orguanidino-aryl substituted alkanoic acid derivatives having thefollowing structural formula

    R.sup.1 --A--(W).sub.a --X--(CH.sub.2).sub.b --(Y)--B--Z--COOR

which are useful for the treatment of thrombosis, apoplexy, cardiacinfarction, inflammation, arteriosclerosis and tumors.

European Patent Application 445,796 A2 discloses acetic acid derivativeshaving the formula

    H.sub.2 N(NH)C--X--Y--CO--Z--CH(Q.sup.1)COOQ.sup.2         (Formula A)

where

Q1 stands for hydrogen, methyl or phenyl,

Q² stands for hydrogen, phenyl-low-alkyl or low alkyl that can becleaved under physiological conditions,

X stands for 1,4-phenylene, 2,5- or 3,6-pyridylene or,1,4-piperidinylene, which is bonded to group Y through the C atom in the4-position,

Y is a group having the formula ##STR6## where Q³ stands for hydrogen,methyl, phenyl, --COOH, --COO-low-alkyl, --CONH(CH₂)₂ --COOH or--CONH(CH₂)₂ --COO-low-alkyl,

Q⁴ hydrogen, methyl or phenyl,

Z a 1,4-piperazinylene group, a 1,4-piperazinylene group which is bondedto the CO group through the N atom in the 1-position or a group havingthe formula

    --NHCH(R.sup.1)-- or --NHCH(COR.sup.2)--

where

R¹ stands for hydrogen, methyl, phenyl or a --COO-low-alkyl,

R² stands for the residue of an α-aminocarboxylic acid bonded throughthe amino group or of an ester or amide thereof, or a group having theformula --NHCH₂ CH₂ --Ar, or --CO--R², or, if applicable, a mono- ordi-low-alkylated carbamoyl group or a pyrrolidinoyl or piperidinoylgroup,

Ar stands for a phenyl or a phenyl substituted by low alkyl, low alkoxy,--COOH, --COO-low-alkyl, --O(CH₂)₁₋₄ --COOH, --O(CH₂)₁₋₄--COO-low-alkyl, --CONH₂, --CONH-low-alkyl, --CON(low alkyl)₂,pyrrolidinoyl or piperidinoyl which are said to have inhibitory actionon the bonding of adhesive proteins to blood platelets as well as bloodplatelet aggregation and cell-cell adhesion.

Pharmazie 29, H.5 (1974) Walsmann, et al., "Synthetische Inhibitoren vonSerinproteinasen" disclosed amidinophenylalkyl/alkenylacid derivatives.

SUMMARY OF THE INVENTION

The present invention relates to a class of compounds represented by theformula ##STR7## or a pharmaceutically acceptable salt thereof, wherein

R₃ and R₄ are each independently selected from the group consisting ofhydrido, alkyl having 1 to 6 carbon atoms, hydroxy, alkoxy having 1 to 6carbon atoms and halo;

X is --CH₂ CH₂ --, --CH═CH--, --C.tbd.C-- or HNCO;

m is an integer from 1 to 3; and

A is the group ##STR8## wherein R₁ is hydrido; hydroxy; alkyl having 1to 6 carbon atoms; alkenyl having 2 to 6 carbon atoms which may beoptionally substituted by halo; alkynyl having 2 to 6 carbon atoms;alkoxycarbonylalkyl; phenylsulfonylalkyl; alkylsulfonylalkyl; phenylwhich may be optionally substituted by alkyl having 1 to 6 carbon atoms,alkoxy having 1 to 6 carbon atoms and hydroxy; a fully unsaturatedheteromonocyclic ring structure having 5 or 6 ring carbon atoms whereinone of the ring carbon atoms is replaced by nitrogen, oxygen or sulfur;or cycloalkyl having 3 to 6 carbon atoms; and

R₂ is hydrido or alkyl having 1 to 6 carbon atoms.

The invention further relates to pharmaceutical compositions comprisinga compound of formula I. Such compounds and compositions have usefulnessas inhibitors of platelet aggregation. The invention also relates to amethod of inhibiting platelet aggregation in a mammal in need of suchtreatment.

A preferred embodiment of the present invention is a compound of theformula ##STR9## R₃ and R₄ are each independently selected from thegroup consisting of hydrido, alkyl having 1 to 6 carbon atoms, hydroxy,alkoxy having 1 to 6 carbon atoms and halo;

X is --CH₂ CH₂ --, --CH═CH--, --C.tbd.C-- or HNCO;

m is an integer from 1 to 3; and

A is the group ##STR10## wherein R₁ is hydrido; hydroxy; alkyl having 1to 6 carbon atoms; alkenyl having 2 to 6 carbon atoms which may beoptionally substituted by halo; alkynyl having 2 to 6 carbon atoms;alkoxycarbonylalkyl; phenylsulfonylalkyl; alkylsulfonylalkyl; phenylwhich may be optionally substituted by alkyl having 1 to 6 carbon atoms,alkoxy having 1 to 6 carbon atoms and hydroxy; a fully unsaturatedheteromonocyclic ring structure having 5 or 6 ring carbon atoms whereinone of the ring carbon atoms is replaced by nitrogen, oxygen or sulfur;or cycloalkyl having 3 to 6 carbon atoms.

Exemplifying this embodiment are the following compounds:

(±)-cis-6- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4-phenyl-2H-pyran-2-one,trifluoroacetate;

(±)-cis-6- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4-hydroxy-2H-pyran-2-one,trifluoroacetate;

(±)-trans-6- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4-hydroxy-2H-pyran-2-one,trifluoroacetate;

(±)-6- 4- 4-(aminoiminomethyl)phenyl!butyl!tetrahydro-2H-pyran-2-one,trifluoroacetate;

(±)-cis-6- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4-methyl-2H-pyran-2-one,trifluoroacetate;

(±)-cis-6- 4-4-(aminoiminomethyl)phenyl!butyl!-4-ethenyltetrahydro-2H-pyran-2-one,trifluoroacetate;

(±)-cis-methyl 2- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-6-oxo-2H-pyran-4-acetate;

(±)-cis-6- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4-(3-pyridinyl)-2H-pyran-2-one,ditrifluoroacetate;

(±)-cis-6- 4- 4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4-(phenylsulfonyl)methyl!-2H-pyran-2-one, trifluoroacetate;

(±)-trans-6- 4-4-(aminiminomethyl)phenyl!butyl!tetrahydro-4-phenyl-2H-pyran-2-one,trifluoroacetate; and

N- 4-(aminoiminomethyl)phenyl!tetrahydro-6-oxo-2H-pyran-2-propanamide,trifluoroacetate.

As used herein, the term "hydrido" denotes a single hydrogen atom (H).This hydrido group may be attached, for example, to a oxygen atom toform a hydroxyl group; or, as another example, two hydrido groups may beattached to a carbon atom to form a --CH₂ -- group.

As used herein, the term "alkyl" either alone or within other terms suchas "alkylcarboxyalkyl" embraces a linear or branched chain saturatedhydrocarbon radical having 1 to 6 carbon atoms. Illustrative of suchradicals are methyl, ethyl, propyl, 1-methylethyl, butyl,2-methylpropyl, 1-methylpropyl, 1,1-dimethylethyl, pentyl,3-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 2,2-dimethylpropyl,1,1-dimethylpropyl, hexyl, and 4-methylpentyl.

As used herein, the term "alkoxy" embraces linear or branchedoxy-containing radicals each having alkyl portions of 1 to 6 carbonatoms. Illustrative of such groups are methoxy, ethoxy, propoxy, butoxy,1-methylethoxy, 2-methylpropoxy, 1-methylpropoxy, 1,1-dimethylethoxy,pentenyloxy, 3-methylbutoxy, 1-methylbutoxy, 1-ethylpropoxy,2-2-dimethylpropoxy, 1,1-dimethylpropoxy, hexenyloxy, and4-methylpentoxy.

As used herein the term "alkenyl" embraces linear or branchedunsaturated hydrocarbon radicals having 2 to 6 carbon atoms andcontaining at least one carbon to carbon double bond, which carbon tocarbon double bond may have either cis or trans geometry within thealkenyl moiety. Said alkenyl moiety may be further substituted with oneor more halo atoms, such as fluoro, chloro or bromo, to provide ahaloalkenyl group. Illustrative of such groups are ethenyl, propenyl,butenyl, isobutenyl, pentenyl, 3-methyl-1-butenyl, 2-methyl-2-butenyl,2,3-dimethyl-2-butenyl and hexenyl.

As used herein the term "alkynyl" embraces linear or branchedunsaturated hydrocarbon radicals having 2 to 6 carbon atoms andcontaining one carbon to carbon triple bond. Illustrative of suchradicals are ethynyl, propynyl, butynyl, pentynyl, and hexynyl.

As used herein the term "halo" embraces halogen atoms. Illustrative ofsuch atoms are chloro (Cl), fluoro (F), bromo (Br) and iodo (I).

As used herein, the term "alkoxycarbonylalkyl" represents the radical ofthe formula

    RCOOR

wherein the R represents an alkyl group having 1 to 6 carbon atoms whichmay or may not be the same. Illustrative of such groups aremethoxycarbonylmethyl and ethoxycarbonylmethyl.

As used herein the term "heteromonocyclic" embraces fully unsaturated,cyclic hydrocarbon radicals having 5 or 6 ring carbon atoms wherein 1 ofthe ring carbons is replaced by nitrogen, oxygen or sulfur. Illustrativeof such radicals are pyridinyl, pyrrolyl, thiophenyl, furanyl andpyranyl. Attachment of the heteromonocyclic structure to the remainingportion of the molecule represented by formula I may be through a ringcarbon atom of the heteromonocyclic structure.

As used herein the term "cycloalkyl" embraces cyclic radicals havingthree to six carbon atoms. Illustrative of such groups are cyclopropyl,cyclobutyl, cyclopentyl and cyclohexyl.

As used herein the term "phenyl" denotes a monocyclic arene in which onehydrogen atom from a carbon atom of the ring has been removed.

Substitution to said phenyl radical can be to any available ring carbonatom.

As the compounds of the present invention may contain one or moreasymmetric carbon atoms, the invention includes all of the possibleenantiomeric and diastereomeric forms of the compounds of formula I. Thecompounds of Formula I which contain two centers of asymmetry mayproduce four possible stereoisomers designated as the RR, RS, SR and ssenantiomers; all four stereoisomers are considered within the scope ofthis invention. As used herein and in the claims the prefix (±) is usedto specify the racemic nature of a particular compound.

The term "phenylsulfonylalkyl" refers to the compound having thefollowing structure ##STR11## wherein R represents an alkyl group having1 to 6 carbon atoms.

The term "alkylsulfonylalkyl" refers to the compound having thefollowing structure

    --RSO.sub.2 R

wherein R represents an alkyl group having 1 to 6 carbon atoms.

Tautomeric forms are also included as well as pharmaceuticallyacceptable salts of such isomers and tautomers.

Compounds of formula I wherein A is 2H-pyran-2-onyl may underphysiological conditions open to give the open-chain acids andcorresponding salts. All such open-chain salts are meant to be includedwithin the present invention.

In the structures and formulas herein, the bond drawn across a bond ofan aromatic ring can be to any available atom on the aromatic ring.

The term "pharmaceutically acceptable salt" refers to a salt prepared bycontacting a compound of formula (I) with an acid whose anion isgenerally considered suitable for human consumption. Examples ofpharmacologically acceptable salts include the hydrochloride,hydrobromide, hydroiodide, sulfate, phosphate, acetate, propionate,lactate, maleate, malate, succinate, and tartrate salts. All of thesesalts may be prepared by conventional means by reacting, for example,the appropriate acid with the corresponding compound of Formula I.

The compounds of formula I were prepared in a conventional manner usingstandard synthetic methods. In general, these benzamidine/lactones wereprepared from the benzonitrile 2 (scheme A), by a three stepprocedure: 1) H₂ S treatment to form the thioamide, 2) alkylation withiodomethane leading to the thioimidate and 3) treatment of thethioimidate with ammonium acetate.

The lactone portion of the molecule was prepared by a variety of methodsdepending on the particular ring substituent R as outlined in schemes B,C and D. The synthesis of hydroxylactones 6 and 7 and intermediates 3and 8 are outlined in scheme B. Thus the halobenzonitrile was coupled toan omega alkynol via a palladium mediated coupling reaction employingtetrakis(triphenylphosphine) palladium (0) for related conditions see:H. A. Dieck and F. R. Heck, J. Organometallic Chem. 259-263(1975)!.Compounds where X=CH₂ CH₂ were prepared by a selective hydrogenationusing palladium on calcium carbonate. Swern oxidation provided thealdehyde intermediate 3. Treatment of 3 with the dianion of methylacetoacetate followed by sodium borohydride reduction gave a mixture ofanti and syn diol esters (4a,5a) which were separated by columnchromatography. Alternatively, reduction with tetramethylammoniumtriacetoxyborohydride afforded 4a selectively cf. D. A. Evans, K. T.Chapman and E. M. Carreira, J. Am. Chem. Soc. 110, 360-78 (1988)!.Saponification afforded the corresponding dihydroxy acid diasteriomers4b and 5b which were converted to the target compounds according toscheme A as either the zwitterionic amidine/acid or amidine/lactonedepending on the workup procedure (see examples). Finally, lactonization(TFA/1,2-dichloroethane) of 4b and β-elimination of 6 (MsCl/Et₃ N) gavethe unsaturated lactone intermediate 8.

Scheme C outlines the general methods used to synthesize a variety ofsubstituted lactones. Stereoselective 1,4-addition (method 1) of vinylcuprate to 8 cf. W. H. Pirkle and P. E. Adams J. Org. Chem. 45, 4117-21(1980)! followed by saponification and treatment of the crude hydroxyacid with triphenylphosphine and diethylazodicarboxylate (DEAD) cf. O.Mitsunobu Synthesis 1-28 (1981)! gave the cis lactone 9. The finaltarget compound was synthesized according to the protocol in scheme A.

Conjugate addition of methyl phenylsulphonylacetate to 8 (method 2) gavethe trans lactone intermediate 10 which underwent isomerization to thecis lactone 11 under the desulfonylation condition of sodium amalgam inmethanol cf. J. A. Marshal, R. C. Andrews and L. Lebioda J. Org. Chem.52, 2378-88 (1987)!. Phenylsulfone 12 was prepared from 10 viadealkylative decarboxylation with sodium chloride in wet DMSO cf. A. P.Krapcho Synthesis 893-914 (1982)! followed by saponification andMitsunobu inversion. Additionally, trans aryl compounds (e.g. 13) weresynthesized via aryl cuprate addition to 8 (method 3). Elaboration tothe final target compound was carried out as outlined in scheme A.

Methods 4, 5 and 6 utilize the intermediate aldehyde 3. Aldolcondensation (method 4) of 3 with the lithium enolate of acetophenoneafforded a β-hydroxy ketone. Esterification with bromoacetylbromide/pyridine followed by samarium iodide induced cyclization of thebromoester afforded the lactone 14 as a single diastereoisomer cf. G. A.Molander and J. B. Etter, J. Am. chem. Soc. 109, 6556-58 (1987)!.Dehydration with thionyl chloride/pyridine followed by selectivehydrogenation using palladium on calcium carbonate gave the cis phenyllactone 15.

Method 5 outlines the synthesis of methyl lactone 19. Treatment of thealdehyde 3 with an in situ generated methallylzinc reagent cf. T. Shono,M. Ishifune and S. Kashimura Chemistry Letters 449-52 (1990)! affordedthe homoallylic alcohol 16. Ozonolysis, acylation and samarium iodideinduced cyclization provided the lactone 17 as a single diastereomer. Atthis stage the benzonitrile was converted to the benzamidine 18 by thesequence outlined in scheme A. Elimination under acidic conditions(trifluoroacetic anhydride/trifluoroacetic acid) followed byhydrogenation of the unsaturated lactone with 10% palladium on carbonafforded the cis methyl lactone 19.

Method 6 is a variation on method 3 featuring an intramolecularWittig-Horner reaction cf. G. R. Weihe and T. C. McMorris J. Org. Chem.43, 3942 (1978)! in order to append a pyridyl moiety on the lactonering. Aldol condensation affords the β-hydroxy ketone 20. Acylation withdimethylphosphonoacetyl chloride/pyridine followed by treatment withsodium hydride provided the unsaturated lactone 21. Catalytichydrogenation (5% Pd/C) gave a cis lactone, which when treated withexcess lithium bis(trimethylsilyl)amide followed by acidic workupafforded the target benzamidine 22 directly cf. R. T. Boere, R. T.Oakley and R. W. Reed, J. Organometallic Chem. 161-7 (1987)!.

Scheme D outlines the method used for compounds of formula 1 whereX=HNCO. Known lactone 23 cf. A. Ijima, H. Mizuno and K. Takahashi Chem.Pharm. Bull. 19, 1053-5 (1971)! was oxidized to the carboxylic acid(NaIO₄ /RuCl₃), converted to the acid chloride (COCl)₂ /DMF (cat.)! andcoupled directly with 4-aminobenzonitrile affording the intermediate 24.Synthesis of the benzamidine according to the procedure outlined inscheme A yielded the desired target compounds.

Scheme E illustrates the synthesis of the chiral hydroxylactone 29 bycombining some synthetic concepts outlined in schemes B and D. The knownhydroxy ester (25), synthesized in 98% e.e., D. F. Taber, L. J.Silverberg Tetrahedron Letters 32, 4227-4230 (1991)! underwent a Claisencondensation with excess lithio t-butyl acetate. Stereoselectivereduction of the hydroxyketoester 26 with tetramethylammoniumtriacetoxyborohydride gave the anti diol ester 27. Hydrolysis,lactonization and silyl protection gave the lactone 28 in high yield.Oxidative cleavage of the trisubstituted olefin and activation of theresulting carboxylic acid to the acid chloride under neutral conditionsS. E. Kelly, T. G. LaCour Synthetic Comm. 22, 859-869 (1992).! followedby coupling with aminobenzamidine dihydrochloride gave the penultimateproduct. Removal of the silyl protecting group using aq. HF inacetonitrile afforded the desired lactone 29. ##STR12##

This invention also relates to a method of inhibiting plateletaggregation and more specifically, a method of treatment involving theadministration of compounds of Formula I to achieve such inhibition.

For the inhibition of platelet aggregation compounds of Formula I may beadministered orally, parenterally, or by inhalation spray or rectally indosage unit formulations containing conventional non-toxicpharmaceutically acceptable carriers, adjuvants and vehicles. The termparenteral as used herein includes, for example, subcutaneous,intravenous, intramuscular, intrasternal, infusion techniques orintraperitonally.

The compounds of the present invention may be administered by anysuitable route, preferably in the form of a pharmaceutical compositionadapted to such a route, and in a dose effective for the treatmentintended. Therapeutically effective doses of the compounds of thepresent invention required to prevent or arrest the progress of themedical condition are readily ascertained by one of ordinary skill inthe art.

Accordingly, the invention provides a class of novel pharmaceuticalcompositions comprising one or more compounds of the present inventionin association with one or more non-toxic, pharmaceutically acceptablecarriers and/or diluents and/or adjuvants (collectively referred toherein as "carrier" materials) and if desired other active ingredients.

The dosage regimen for treating a condition with the compounds and/orcompositions of this invention is based on a variety of factors,including the type, age, weight, sex and medical condition of thepatient; the severity of the condition; the route of administration; andthe particular compound employed. Thus dosage regimen may vary widely.Dosage levels of the order from about 0.01 mg to about 150 mg perkilogram of body weight per day are useful in the treatment of theabove-indicated conditions (from about 10 mg to about 10500 mg perpatient per day). For oral administration a daily dose of from about0.01 to 150 mg/Kg body weight, particularly from about 1 to 30 mg/Kgbody weight may be appropriate. For administration by injection apreferred daily dose would be from about 0.01 to 50 mg/Kg body weight.

For oral administration, the pharmaceutical composition may be in theform of, for example, a tablet, capsule, suspension or liquid. Thepharmaceutical composition is preferably made in the form of a dosageunit containing a particular amount of the active ingredient. Examplesof such dosage units are tablets or capsules. These may contain, forexample, an amount of active ingredient from about 1 to 250 mg,preferably from about 25 to 150 mg. A suitable daily dose for a mammalmay vary widely depending on the condition of the patient and otherfactors.

The active ingredient may also be administered by injection as acomposition wherein, for example, saline, dextrose or water may be usedas a suitable carrier. A suitable daily dose would typically be about0.01 to 50 mg/kg body weight injected per day in multiple dosesdepending on the condition being treated.

For administration, the compounds of this invention are ordinarilycombined with one or more adjuvants appropriate to the indicated routeof administration. The compounds may be admixed with lactose, sucrose,starch powder, cellulose esters of alkanoic acids, cellulose alkylesters, talc, stearic acid, magnesium stearate, magnesium oxide, sodiumand calcium salts of phosphoric and sulphuric acids, gelatin, acacia,sodium alginate, polyvinylpyrrolidone, and/or polyvinyl alcohol, andtableted or encapsulated for convenient administration. Alternatively,the compounds may be dissolved in water, polyethylene glycol, propyleneglycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil,benzyl alcohol, sodium chloride, and/or various buffers. Other adjuvantsand modes of administration are well and widely known in thepharmaceutical art.

The pharmaceutical compositions may be made up in a solid form such asgranules, powders or suppositories or in a liquid form such assolutions, suspensions or emulsions. The pharmaceutical compositions maybe subjected to conventional pharmaceutical operations such assterilization and/or may contain conventional pharmaceutical adjuvantssuch as preservatives, stabilizers, wetting agents, emulsifiers,buffers, etc.

The following Examples are intended to further illustrate the presentinvention and not to limit the invention in spirit or scope. In theExamples temperature is in degrees Celsius unless otherwise expresslyset forth.

EXAMPLE 1

Preparation of (±)-4-(Aminoiminomethyl)-βS, δR-dihydroxybenzenenonamoicacid and (±)-cis-6- 4-4-Aminoiminomethyl)phenyl!butyl!tetrahydro-4-hydroxy-2H-pyran-2-one,trifluoroacetate ##STR13## A. Preparation of5-(4-cyanophenyl)-4-pentynol

To a solution of 4-bromobenzonitrile (105.0 g, 0.577 mo!) andtriethylamine (108.1 g, 1.07 mo!) in 900 mL of acetonitrile undernitrogen was added 4-pentynol (50.0 g, 0.594 mol) dissolved inacetonitrile followed by tetrakis(triphenylphosphine)palladium(0) (5.00g, 3.23 mmol). The reaction flask was wrapped in aluminum foil and themixture was refluxed for 20 hours, cooled to room temperature andfiltered. The filtercake was washed with acetonitrile and the filtratewas evaporated under reduced pressure. The residue was dissolved inEtOAc and washed successively with water, 5% aq. HCl, water, 5% aq. K₂CO₃, water, and brine. The organic layer was dried (MgSO₄) and thesolvent removed under reduced pressure. The residue was dissolved in 1.8L of diethyl ether, treated with charcoal, filtered and the filtratereduced to a volume of 550 mL on a steam bath. A solid yellowprecipitate formed upon cooling to -30° C. The solid was filtered andwashed with cold diethyl ether and dried affording 72.90 g of the titleproduct (m.p. 76.83° C.) The filtrate was concentrated under reducedpressure and chromatographed (EtOAc: hexane, 1:1) affording 10.2 g oftitle product after recrystallization from diethyl ether (m.p. 81°-84°C., 78% combined yield).

Anal. calc'd. for C₁₂ H₁₁ NO: C, 77.82; H, 5.99; N, 7.56. Found: C,77.30; H, 6.08; N, 7.38.

B. Preparation of 5-(4-cyanophenyl)pentanol

To a solution of the product of step A (65.0 g, 0.351 mol) in methanol(1 L) was added 5% Pd/CaCO₃ (6.50 g) and hydrogenated at 5 psi for 3.5hours. The catalyst was removed and the solvent evaporated under reducedpressure. The light brown residue was dissolved in CH₂ Cl₂ (150 mL) andpassed through a bed of silica gel and eluted with CH₂ Cl₂. The solventwas removed under reduced pressure yielding 55.6 g (84%) of the titleproduct which solidified on standing (m.p. 39°-41° C.).

Anal. calc'd. for C₁₂ H₁₅ NO: C, 76.16; H, 7.99; N, 7.40. Found: C,76.09; H, 7.93; N, 7.28.

C. Preparation of 5-(4-cyanophenyl)pentanal

A solution of DMSO (9.36 g, 120 mmol) in 20 mL of CH₂ Cl₂ was addeddropwise over 10 minutes to a solution of oxalyl chloride (10.16 g, 80mmol) in CH₂ Cl₂ (80 mL) at -70° C. under an atmosphere of nitrogen.After stirring for 15 minutes at -70° C. a solution of the product ofstep B (7.56 g, 40.0 mmol) in CH₂ Cl₂ (50 mL) was added dropwise over 10minutes and the solution was stirred at -70° C. for 15 minutes. Neattriethylamine (24.2 g, 240 mmol) was added rapidly, the ice bath wasremoved and the mixture allowed to warm to ambient temperature over 30minutes. The reaction mixture was diluted with CH₂ Cl₂ (100 mL), washedsuccessively with water, 1N NaHSO₄ and brine, dried (MgS0₄), filteredthrough a pad of silica gel and eluted with CH₂ Cl₂. The solvent wasevaporated under reduced pressure affording 7.23 g of title product(97%) as a thick oil of sufficient purity to be used in the nextreaction.

¹ H-NMR (300 MHz, CDCl₃) δ1.67 (m, 4H), 2.47 (m, 2H), 2.70 (m, 2H), 7.37(d, J=8 HZ, 2H), 7.48 (d, J=8 Hz, 2H), 9.77 (s, 1H).

D. Preparation of methyl 9-(4-cyanophenyl)-3,5-dihydroxynonanoate Neatmethyl acetoacetate (2.33 g, 20 mmol) was added dropwise to a suspensionof NaH (60% oil suspension) (800 mg, 20.0 mmol) in dry THF (40 mL) at 0°C. under a nitrogen atmosphere. The resulting solution was stirred at 0°C. for 15 minutes and treated with a 1.6M solution (12.5 mL, 20 mmol) ofn-BuLi in hexane added dropwise over 5 minutes. The yellow solution wasstirred at 0° C. for 15 minutes whereupon the product of step C (3.74 g,20 mmol) in THF (10 mL) was added via canula and stirred at 0° C. for 30minutes. The reaction mixture was poured into 1N NaHSO₄ (100 mL) andextracted with ETOAc, dried (MgSO₄), filtered and evaporated underreduced pressure. The crude hydroxy-keto ester was dissolved in methanol(50 mL) and cooled to -30° C. Solid NaBH₄ (760 mg, 20 mmol) was addedand the reaction was stirred at -30° C. for 2 hours, poured into 1NNaHSO₄, extracted with EtOAc, dried (MgSO₄), filtered and concentratedunder reduced pressure. Silica gel chromatography (EtOAc:toluene, 4:6)afforded two diastereomeric diol esters.

Diastereomer A (610 mg) solidified to a waxy solid on standing.

Anal. calc'd. for C₁₇ H₂₃ NO₄ : C, 66.86; H, 7.59; N, 4.59. Found: C,66.16; H, 7.74; N, 4.35.

¹ H-NMR (300 MHz, CDCl₃ /D₂ O) δ1.25-1.75 (m, 8H), 2.50 (d, J=6 Hz, 2H),2.68 (t, J=7 Hz, 2H), 3.72 (s, 3H), 3.86 (m, 1H), 4.26 (pent., J=7 Hz,1H), 7.27 (d, J=8 Hz, 2H), 7.57 (d, J=8 Hz, 2H) .

Diastereomer B was recrystallized from methyl t-butyl ether/diisopropylether (1:1) affording 720 mg of product (m.p. 88.93° C. ) .

Anal. calc'd. for C₁₇ O₂₃ NO₄ : C, 66.86; H, 7.59; N, 4.59. Found: C,66.65; H, 7.72; N, 4.51.

¹ H-NMR (300 MHz, CDCl₃ /D₂ O) δ1.3-1.7 (m, 8H), 2.47 (dd, J=5 Hz, J=12Hz, 1H), 2.55 (dd, J=7 Hz, J=12 Hz, 1H), 2.68 (t, J=7 Hz, 2H), 3.72 (s,3H), 3.90 (m, 1H), 4.34 (m, J=5 Hz, 1H), 7.27 (d, J=8 Hz, 2H), 7.57 (d,J=8 Hz, 2H).

The syn/ anti assignment of the two diol diastereomers was determined byNMR analysis of the corresponding lactones as compared to the literaturefor NMR analysis of related compounds see: W. F. Hoffman, et al. J. Med.Chem. 29, 159-169 (1986)! and is described below.

E. Preparation of trans-6-4-(4-Cyanophenyl)butyl!3,4,5,6-tetrahydro-4-hydroxy-2H-pyran-2-one

To a solution of diasteriomer A, step D (55 mg, 0.180 mmol) in THF (3mL) was added 1N aq. NaOH (200 uL, 0.20 mmol) and the solution stirredat room temperature for 30 minutes. The reaction mixture was acidifiedby adding 2N HCl (200 uL), stirred for 5 minutes, treated with MgSO₄,filtered and concentrated under reduced pressure. The residue was takenup in 1,2-dichloroethane (5 mL). Trifluoroacetic acid (20 uL, 0.26 mmol)was added and the solution stirred at 50° C. for 1 hour. Removal of thesolvent under reduced pressure gave 50 mg of product as a colorless oil.¹ H-NMR (300 MHz, CDCl₃) δ1.30-1.80 (m, 7H), 1.90-2.00 (m, 1H),2.40-2.60 (m, 4H), 4.40 (m, 1H), 4.70 (m, 1H), 7.27 (d, J=8 Hz, 2H),7.57 (d, J=8 Hz, 2H).

F. Preparation of cis-6-4-(4-Cyanophenyl)butyl!-3,4,5,6-tetrahydro-4-hydroxy-2H-pyran-2-one

The title compound was synthesized from diasteriomer B, step D (55 mg,0.180 mmol) in a manner similar to step E affording 50 mg of a colorlessoil.

¹ H-NMR (300 MHz, CDCl₃) δ1.40-1.80 (m, 7H), 2.19-2.30 (m, 1H), 2.48(dd, J=8 Hz, J=16 Hz, 1H), 2.78 (t, J=7 Hz, 2H), 2.91 (dd, J=6 Hz, J=16Hz, 1H), 4.15-4.32 (m, 2H), 7.27 (d, J=8 Hz, 2H), 7.57 (d, J=8 Hz, 2H).

The stereochemical assignment was based on the observation that theaxial hydroxyl group in the lactone of diastereomer A deshields the C-6lactone proton (δ4.70) by 0.47 ppm compared to the chemical shiftassignment of the corresponding proton on the lactone diastereomer B(δ˜4.2) (see W. F. Hoffman, et a!., 1986). Additional supportingevidence was obtained when the hydroxy-keto ester intermediate in step Dwas stereoselectively reduced with tetramethylammoniumtriacetoxyborohydride cf. D. A. Evans, K. T. Chapman and E. M. CarreiraJ. Am. Chem. Soc. 110, 3560-78 (1988)! to give the anti diol ester whoseNMR spectrum was identical in every respect to diastereomer B in step D.

G. Preparation of (±)-4-(Aminoiminomethyl)-βS, R-dihydroxybenzenonamoicacid and (±)-cis-6-(4-(4-(aminoiminomethyl)phenyl)butyl!-3,4,5,6-tetrahydro-4-hydroxy-2H-pyran-2-one

To a solution of diasteriomer B, step D (392 mg, 1.29 mmol) in MeOH (10mL) was added aq. 1N NaOH (1.9 mL, 1.9 mmol) and the solution stirred atroom temperature for 1 hour. The reaction was poured into 1N NaHSO₄,extracted with EtOAc, dried (MgSO₄) and evaporated under reducedpressure. To the crude dihydroxy acid in pyridine (10 mL) was addedtriethylamine (781 mg, 7.74 mmol). Hydrogen sulfide gas was bubbledthrough the solution for 5 minutes at room temperature, then stopperedand stirred at room temperature for 2 days. The solvent was removedunder reduced pressure and the yellow residue partitioned between EtOAcand 1N NaHSO₄. The organic layer was dried (MgSO₄), filtered andevaporated under reduced pressure. To the resulting thioamide in acetone(10 mL) was added iodomethane (1.83 g, 12.90 mmol) and the reactionstirred at 55° C. under an atmosphere of nitrogen for 35 minutes.Removal of the solvent under reduced pressure afforded the crudethioimidate HI. To this yellow residue was added anhydrous ammoniumacetate (198 mg, 2.58 mmol) and methanol (10 mL). The solution wasstirred at 55° C. for 3.5 hours under an atmosphere of nitrogen thenconcentrated under reduced pressure. The residue was dissolved in water(2 mL) and the product was precipitated as the zwitterion by theaddition of acetone (30 mL). The precipitate was filtered and washedsuccessively with water:acetone (1:15), acetone, acetonitrile, anddiethyl ether affording 130 mg of the title compound (acid) as anoff-white amorphous product (m.p. 238°-242° C.)

Anal. calc'd. for C₁₆ H₂₄ N₂ O₄ O.4H₂ O: C, 60.55; H, 7.92; N, 8.88.Found: C, 60.92,; H, 7.70; N, 8.46.

The filtrate was evaporated under reduced pressure and purified byreverse phase chromatography on a Waters® C-18 Delta Pak column using a0.05% TFA/water:acetonitrile gradient affording 210 mg of the titlecompound (lactone) as the TFA salt after dissolving the residue in hotCH₃ CN, cooling, then precipitation with Et₂ O (m.p. 150°-152° C. dec.).

Anal. calc'd. for C₁₈ H₂₃ N₂ O₅ F₃ O.5H₂ O: C, 52.29; H, 5.85; N, 6.78.Found: C, 52.16; H, 5.62; N, 6.71.

EXAMPLE 2

Preparation of (±)-trans-6- 4-4-(Aminoiminomethyl)phenyl!butyl!tetrahydro-4-hydroxy-2H-pyran-2-one,trifluoroacetate ##STR14##

The title compound was prepared from the product of example 1, step D(diasteriomer A, 350 mg, 1.15 mmol) in a manner similar to example 1,step G affording 115 mg (25%) of lactone product as the TFA salt afterreverse phase chromatography (m.p. 159°-160° C.).

Anal. calc'd. for C₁₈ H₂₃ N₂ O₅ F₃ O.2H₂ O: C, 52.99; H, 5.78; N, 6.87.Found: C, 52.90; H, 5.73; N, 6.85.

EXAMPLE 3

Preparation of (±)-ethyl 4-(aminoiminomethyl)-βS,δR-dihydroxybenzeneonanoate ##STR15## A. Preparation ofanti-ethyl-9-(4-cyanophenyl)-3,5-dihydroxynonanoate

The title compound was prepared from the product of example 1, step C(1.87 g, 10 mmol) in a manner similar to example 1, step D substitutingethyl acetoacetate (1.30 g, 10 mmol) in place of methyl acetoacetate.The intermediate hydroxy-keto ester was purified by silica gelchromatography (EtOAc:hexane 1:1) giving 2.54 g of partionally purifiedproduct. The carbonyl was stereoselectively reduced usingtetramethylammonium triacetoxyborohydride (D. A. Evans, 1988). Thus,tetramethylammonium triacetoxyborohydride (16.6 g, 63.12 mmol) wasdissolved in 1:1 acetonitrile: acetic acid (40 mL) and stirred at roomtemperature for 30 minutes, then cooled to -40° C. The hydroxy-ketoester (2.5 g, 7.89 mmol) in acetonitrile (10 mL) and the solutionstirred at -40° C. for 2 hours then at -10° C. for 3 hours. The reactionwas quenched by the addition of sat'd aq. Na/K tartrate (20 mL) thenpoured into sat'd aq. NaHCO₃ (200 mL). Solid NaHCO₃ was added carefullyuntil gas evolution ceased then extracted (2×) with EtOAc, dried(MgSO₄), filtered and evaporated under reduced pressure. Silica gelchromatography (EtOAc:hexane 6:4) afforded 1.53 g (48% overall). Ananalytical sample was prepared by crystallizing from diisopropyl ether(m.p. 86.5°-87.5° C.).

Anal. calc'd. for C₁₈ H₂₅ NO₄ : C, 67.69; H, 7.89; N, 4.39. Found: C,67.79; H, 8.14; N, 4.37.

¹ H-NMR (300 MHz, CDCl₃) δ1.28 (t, J=7 Hz, 3 HN), 1.30-1.73 (m, 8 HN),2.45 (dd, J=5 Hz, J=13 Hz, 1H), 2.53 (dd, J=7 Hz, J=13 Hz, 1H), 2.68 (t,J=7 Hz, 2H), 3.91 (m, 1H), 4.18 (q, J=7 Hz, 2H), 4.36 (m, 1H), 7.27 (d,J=8 Hz, 2H), 7.57 (d, J=8 Hz, 2H).

B. Preparation of (±)-ethyl 4-(aminoiminomethyl)-βS,δR-dihydroxybenzenenonanoate

The title compound was prepared from the product of step A (1.50 g, 4.70mmol) in a manner similar to example 1, step G affording 830 mg (39%) ofproduct as the TFA salt after reverse phase chromatography andtrituration of the product with acetonitrile (m.p. 158°-160° C.).

Anal. calc'd. for C₂₀ H₂₉ N₂ O₆ F₃ 0.25H₂ O: C, 52.80; H, 6.54; N, 6.16.Found: C, 52.70; H, 6.36; N, 6.29.

EXAMPLE 4

Preparation of (±)-cis-6-4-(4-(Aminoiminomethyl)phenyl)butyl!-4-ethenyl-tetrahydro-2H-pyran-2-one,trifluoroacetate ##STR16## A. Preparation of 6-4-(4-cyanophenyl)butyl!-5,6-dihydro-2H-pyran-2-one

To a solution of the product of example 1, step F (2.22 g, 8.13 mmol)and triethylamine (2.06 g, 20.40 mmol) in CH₂ Cl₂ (40 mL) at 0° C. wasadded methanesulfonyl chloride (930 mg, 8.13 mmol). The reaction wasstirred for 10 minutes at 0° C. then at ambient temperature for 2.5hours. The reaction mixture was concentrated under reduced pressure,partitioned between EtOAc and 1N NaHSO₄, then washed successively with10% K₂ CO₃, brine and dried (MgSO₄). Evaporation of the solvent andsilica gel chromatography (EtOAc:hexane 1:1) afforded 608 mg (30%) ofproduct (m.p. 66°-69° C.).

Anal. calc'd. for C₁₆ N₁₇ NO₂ : C, 75.27; H, 6.71; N, 5.49. Found: C,75.00; H, 6.79; N, 5.46.

B. Preparation of trans-6-4-(4-cyanophenyl)butyl)-4-ethenyl-3,4,5,6-tetrahydro-2H-pyran-2-one

To a stirred solution of vinyltributyltin (7.45 g, 23.52 mmol) in dryTHF (15 mL) at -78° C. under an atmosphere of nitrogen was addeddropwise a 1.6M solution of nBuLi (14.7 mL, 23.52 mmol) in hexane. After10 minutes at -78° C., solid CuCN was added all at once under a streamof nitrogen. The mixture was slowly warmed to -30° C. over 45 minutesduring which time the CuCN dissolved. The resulting solution was cooledto -60° C. and the product of step A (1.50 g, 5.88 mmol) in THF (10 mL)was added. The solution was warmed to -20° C. over 30 minutes thenpoured into a rapidly stirred solution of dilute aq. NH₄ OH/NH₄ Cl andstirred for 15 minutes. The aqueous phase was extracted with Et₂ O,dried (MgSO₄), filtered and evaporated under reduced pressure. Thetetrabutyltin biproduct was simply removed by stirring the residue withhexane and carefully decanting the solvent (repeated 2×), affording 1.52g (92%) of title product of sufficient purity to be used in the nextreaction.

¹ N-NMR (300 MHz, CDCl₃) δ1.25-1.85 (m, 8H), 2.47 (dd, J=7 Hz, J=17 Hz,1H), 2.60 (dd, J=5 Hz, J=17 Hz, 1H), 2.68 (t, J=7 Hz, 2H), 2.76 (m, 1H),4.34 (m, 1H), 5.02-5.18 (m, 2H), 5.75-5.89 (m, 1H), 7.27 (d, J=8 Hz,2H), 7.57 (d, J=8 Hz, 2H).

C. Preparation of cis-6-4-(4-cyanophenyl)butyl!-4ethenyl-3,4,5,6-tetrahydro-2H-pyran-2-one

To a solution of the product of step B (1.52 g, 5.47 mmol) in 15 mL ofMeOH:THF (2:1) was added 1H NaOH (5.9 mL, 5.9 mmol) and the reactionstirred at room temperature for 1.5 hours. The reaction was diluted withwater and washed with Et₂ O. The aqueous phase was acidified with 1HNaHSO₄, extracted with EtOAc, dried (MgSO₄), filtered and evaporatedunder reduced pressure. The residue and Ph₃ P (1.86 g, 7.11 mmol) wasimmediately dissolved in dry THF (20 mL) and cooled to -40° C. Neatdiethyl azodicarboxylate (1.27 g, 7.42 mmol) was added dropwise viasyringe and the solution stirred at -40° C. for 3 hours. Concentrationof the solvent under reduced pressure and silica gel chromatography(EtOAc:hexane 4:6) of the residue afforded 1.05 g (69%) of the titleproduct as a colorless oil. NMR spectrum analysis indicated the productwas contaminated with ˜10% of the trans isomer.

¹ H-NMR (300 MHz, CDCl₃) δ1.25-1.80 (m, 7H), 1.90-2.00 (m, 1H), 2.24(dd, J=11 Hz, J=17 Hz, 1H), 2.55-2.79 (m, 4H), 4.30 (m, 1H) 5.02-5.14(m, 2H), 5.66-5.80 (m, 1H), 7.27 (d, J=8 Hz, 2H), 7.57 (d, J=8 Hz, 2H).

D. Preparation of (±)-cis-6-4-(4-(aminoiminomethyl)phenyl)butyl!-4-ethenyl-tetrahydro-2H-pyran-2-one

The title compound was prepared from the product of step C (1.00 g, 3.53mmol) in a manner similar to example 1 step G with a modification in thepurification. The residue was dissolved in acetonitrile and TFA (402 mg,3.53 mmol) was added. The solvent was evaporated under reduced pressureand the residue redissolved in a minimal amount of acetonitrile.Addition of Et₂ O caused the precipitation of the crude product whichwas filtered and washed with Et₂ O. The material was briefly suspendedin ice water, filtered, washed with ice water and dried.Recrystallization from acetonitrile afforded 720 mg (50%) of product asthe TFA salt (m.p. 186°-187° C. dec.).

Anal. calc'd. for C₂₀ N₂₅ N₂ O₄ F₃ 0.3H₂ O: C, 57.14; H, 6.15; N, 6.66.Found: C, 57.17; H, 6.30; N, 6.59.

EXAMPLE 5

Preparation of (±)-cis-methyl 2- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-6-oxo-2H-pyran-4-acetate##STR17## A. Preparation of trans-6- 4-(4-cyanophenyl)butyl!-4-2-methoxy-2-oxo-1-(phenylsulphonyl)ethyl!3,4,5,6-tetrahydro-2H-pyran-2-one

To a solution of the product of example 4, step A (2.00 g, 7.83 mmol)and methyl phenylsulphonylacetate (2.01 g, 9.41 mmol) in 15 mL oftBuOH:DMSO (3:8) stirred at room temperature under an atmosphere ofnitrogen was added solid KOtBu (180 mg, 1.61 mmol). The resultingsolution was stirred at room temperature for 18 hours, poured intoEtOAc, washed successively with 1N NaHSO₄, water, brine and dried(MgSO₄). The solvent was removed under reduced pressure and silica gelchromatography (EtOAc:hexane 1:1) of the residue afforded 3.01 g (82%)of the title product as oil. NMR spectrum analysis indicated a 1:1mixture of phenylsulphone diastereomers.

¹ N-NMR (300 MHz, CDCl₃) δ3.00 (m, 1H), 3.50, 3.59(2 s, OMe), 3.97, 4.00(2 d, J=7 Hz, J=9 Hz, 1H, PhSO₂ CHRCO₂ R) , 4.36, 4.47 (2 m, 1H, H₆),7.27 (m, 2H), 7.53-7.67 (m, 4H), 7.73 (t, J=8 Hz, 1H), 7.89 (m, 2H).

B. Preparation of cis-6- 4-(4-cyanophenyl)butyl!-4-(2-methoxy-2-oxo)ethyl!-3,4,5,6-tetrahydro-2H-pyran-2-one

To a solution of the product of step A (2.76 g, 5.88 mmol) in 60 mL ofMeOH:THF (1:1) at room temperature under an atmosphere of nitrogen wasadded solid Na₂ HPO₄ (10.02 g, 70.60 mmol) . To this mechanicallystirred mixture was added powdered 6% Na(Hg) (25.5 g, 66 mmol) in 3portions over 3 hours. The mixture was stirred at room temperature foran additional 2 hours then quenched with aq. 0.4N tartaric acid (200mL). The mixture was diluted with water and extracted with EtOAc (2×),dried (MgSO₄), filtered and evaporated under reduced pressure. Silicagel chromatography of the residue (EtOAc:CH₂ Cl₂ 1:4) afforded 1.40 g(72%) of product as an oil. NMR spectrum analysis indicates a 4:1mixture of cis/trans isomers.

¹ N-NMR (300 MHz, CDCl₃) δ1.15 (m, 1H), 1.35-1.75 (m, 6H), 2.00 (m, 1H),2.13 (dd, J=10 Hz, J=17 Hz, 1H), 2.35 (d, J=7 Hz, 2H), 2.38-2.52 (m,1H), 2.68 (t, J=7 Hz, 2H), 2.77 (dd, J=5 Hz, J=17 Hz, 1H), 3.70 (s, 3H),4.30 (m, 1H), 7.28 (d, J=8 Hz, 2H), 7.57 (d, J=8 Hz, 2H).

C. Preparation of (±)-cis-methyl 2- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-6-oxo-2H-pyran-4-acetate

The title compound was prepared from the product of step B (1.40 g, 4.25mmol) in a manner similar to example 1, step G with a modification inthe purification. The residue was dissolved in acetonitrile and TFA (910mg) was added. The solvent was concentrated under reduced pressure andthe residue redissolved in a minimal amount of acetonitrile. The crudeproduct was precipitated by the addition of Et₂ O, filtered and washedwith Et₂ O. The dry filtercake was washed with ice water and dried.Reprecipitation from CH₃ CN/Et₂ O gave 1.25 g (64%) of title product asa 4:1 mixture of cis/trans isomers (m.p. 161°-165° C.).

Anal. calc'd. for C₂₁ N₂₇ N₂ O₆ F₃ 0.3H₂ O: C, 54.08; H, 5.98; N, 6.01.Found: C, 53.95; H, 5.87; N, 6.01.

EXAMPLE 6

Preparation of (±)-cis-6- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4-(phenylsulfonyl)methyl!-2H-pyran-2-one, trifluoroacetate ##STR18## A.Preparation of trans-6-4-(4-cyanophenyl)butyl!-4-phenylsulfonylmethyl-3,4,5,6-tetrahydro-2H-pyran-2-one

A solution of the product of example 5, step A (2.74 g, 5.84 mmol) andsodium chloride (370 mg, 63 mmol) in 4.5 mL of DMSO (containing 1.6%water) was stirred in a 165° C. oil bath for 1.5 hours then at 180° C.for 30 minutes. The reaction mixture was cooled, diluted with EtOAc andwashed successively with 1N HCl, water, brine and dried (Na₂ SO₄).Evaporation of the solvent under reduced pressure and silica gelchromatography (EtOAc:hexane 1:1) of the residue afforded 1.12 g (47%)of gummy product.

¹ N-NMR (300 MHz, CDCl₃) δ1.35-1.83 (m, 6H) , 1.90-2.10 (m, 2H), 2.38(m, 1H), 2.60-2.80 (m, 4H), 3.02-3.20 (m, 2H), 4.40 (m, 1H), 7.27 (d,J=8 Hz, 2H), 7.55-7.67 (m, 4H), 7.72 (t, J=8 Hz, 1H), 7.92 (d, J=7 Hz,2H).

B. Preparation of cis-6-4-(4-cyanophenyl)butyl!-4-phenylsulfonylmethyl-3,4,5,6-tetrahydro-2H-pyran-2-one

The title compound was prepared from the product of step A (1.18 g, 2.87mmol) in a manner similar to example 4, step C affording 1.00 g (85%) ofproduct after silica gel chromatography (EtOAc:hexane 1:1).

¹ H-NMR (300 MHz, CDCl₃) δ1.35-1.75 (m, 7H), 2.20-2.34 (m, 2H),2.60-2.75 (m, 3H), 2.85 (ddd, J=2 Hz, J=6 Hz, J=17 Hz, 1H), 4.29 (m,1H), 7.27 (d, J=8 Hz, 2H), 7.53-7.65 (m, 4H), 7.90 (t, J=8 Hz, 1H), 7.91(d, J=7 Hz, 2H).

C. Preparation of (±)-cis-6- 4-4-(aminoiminomlethyl)phenyl!butyl!tetrahydro-4-(phenylsulfonyl)methyl!-2H-pyran-2-one, trifluoroacetate

The title compound was prepared from the product of step B (1.00 g, 2.43mmol) in a manner similar to example 1, step G affording 910 mg (69%) ofproduct after reverse phase chromatography and trituration withacetonitrile (m.p. 233.5°-234.5° C.) .

Anal. calc'd. for C₂₅ N₂₉ N₂ O₆ F₃ S: C, 55.34; H, 5.39; N, 5.16; S,5.91. Found: C, 55.23; H, 5.36; N, 5.17; S, 6.09.

EXAMPLE 7

Preparation of (±)-trans-6- 4-4-(Aminoimino)phenyl)butyl!tetrahydro-4-phenyl-2H-pyran-2-one,trifluoroacetate ##STR19## A. Preparation Of trans-6-4-(4-cyanophenyl)butyl!-4-phenyl-3,4,5,6-tetrahydro-2H-pyran-2-one

The title compound was prepared from the product of example 4, step A(384 mg, 1.50 mmol) in a manner similar to example 4, step Bsubstituting phenyllithium for the n-BuLi/vinyltributyltin combinationto give 196 mg (39%) of product after silica gel chromatography (35%EtOAc in hexane ).

¹ N-NMR (300 MHz, CDCl₃) δ1.3-1.85 (m, 6H), 2.04 (m, 2H), 2.67 (t, J=7Hz, 2H), 2.70-2.85 (m, 2H), 3.34 (pent., J=7 Hz, 1H), 4.37 (m, 1H),7.15-7.40 (m, 7H), 7.57 (d, J=8 Hz, 2H).

B. Preparation of (±)-trans-6- 4-4-(Aminoimino)phenyl)butyl!tetrahydro-4-phenyl-2H-pyran-2-one,trifluoroacetate

The title compound was prepared from the product of step A (264 mg, 792mmol) in a manner similar to example 1 step G affording 165 mg (45%) ofproduct as the TFA salt following reverse phase chromatography (m.p.208°-210° C.).

Anal. calc'd. for C₂₄ N₂₇ N₂ O₄ F₃ : C, 62.06; H, 5.86; N, 6.03. Found:C, 62.08; H, 5.93; N, 6.03.

EXAMPLE 8

Preparation of (±)-cis-6- 4-4-(Aminoiminomethyl)phenyl)butyl!tetrahydro-4-phenyl-2H-pyran-2-one,trifluoroacetate ##STR20## A. Preparation of7-(4-cyanophenyl)-3-hydroxy-1-phenylheptan-1one

To a solution of diisopropylamine (1.01 g, 10 mmol) in dry THF (20 mL)at 0° C. under an atmosphere of nitrogen was added a 1.6M solution ofn-BuLi (6.25 mL, 10 mmol) in hexane. After 15 minutes at 0° C. thesolution was cooled to -78° C. and acetophenone (1.2 g, 10 mmol) wasadded dropwise via syringe. After 10 minutes a solution of the productof example 1 step C (1.87 g, 10 mmol) in 10 mL of THF was added viacannula. The solution was stirred at -78° C. for 30 minutes then pouredinto 1N NaHSO₄, extracted with EtOAc, dried (MgSO₄), filtered andevaporated under reduced pressure. Trituration of the residue withhexane afforded 2.54 g (83%) of title product.

¹ H-NMR (300 MHz, CDCl₃) δ1.40-1.75 (m, 6H), 2.70 (t, J=7 Hz, 2H), 3.03(dd, J=9 Hz, J=18 Hz, 1H), 3.16 (dd, J=4 Hz, J=18 Hz, 1H), 4.22 (m, 1H),7.27 (d, J=8 Hz, 2H), 7.40-7.65 (m, 5H), 7.95 (d, J=8 Hz, 2H).

B. Preparation of3-(2-bromo-1-oxoethoxy)-7-(4-cyanophenyl)-1-phenylheptan-1-one

To a solution of the product of step A (2.42 g, 7.88 mmol) and pyridine(7 mL) in Et₂ O at 0° C. was added bromoacetyl bromide (2.37 g, 11.82mmol) . The mixture was stirred for 1 hour at 0° C. then at roomtemperature for 30 minutes. The mixture was diluted with Et₂ O, washedsuccessively with 1N NaHSO₄, sat'd NaHCO₃ and dried (MgSO₄). Evaporationof the solvent under reduced pressure afforded 3.0 g (89%) of titleproduct.

¹ H-NMR (300 MHz, CDCl₃) δ1.35-1.80 (m, 6H), 2.68 (t, J=7 Hz, 2H), 3.12(dd, J=7 Hz, J=18 Hz, 1H), 3.39 (dd, J=7 Hz, J=18 Hz, 1H), 3.76 (s, 2H),5.49 (pent., J=7 Hz, 1H), 7.26 (d, J=8 Hz, 2H), 7.49 (t, J=8 Hz, 1H),7.52-7.63 (m, 4H), 7.94 (d, J=8 Hz, 2H).

C. Preparation of rel-(4R, 6R)-6-4-(4-cyanophenyl)butyl!-4-hydroxy-4-phenyl-3,4,5,6-tetrahydro-2H-pyran-2-one

To a solution of the product of step B (2.71 g, 6.35 mmol) in dry THF(50 mL) at -78° C. under an atmosphere of nitrogen was added a 0.1Msolution of SmI₂ (135 mL, 13.5 mmol) over 10 minutes. The blue solutionwas stirred at -78° C. for 1 hour, poured into dilute aq. HCl, extractedwith EtOAc, washed with sat'd NaHCO₃, dried (MgSO₄), filtered, and thesolvent evaporated under reduced pressure. Silica gel chromatography(35-50% EtOAc/hexane) afforded 1.90 g (86%) of title product.

¹ H-NMR (300 MHz, CDCl₃) δ1.40-1.90 (m, 6H), 1.99 (dd, J=13 Hz, J=15 Hz,1H), 2.11 (dd, J=2 Hz, J=15 Hz, 1H), 2.60 (t, J=7 Hz, 2H), 2.90 (s, 2H),4.83 (m, 1H), 7.27 (d, J=8 Hz, 2H), 7.30-7.47 (m, 5H), 7.57 (d, J=8 Hz,2H).

D. Preparation of cis-6-4-(4-cyanophenyl)butyl!-4-phenyl-3,4,5,6-tetrahydro-2H-pyran-2-one

To a solution of the product of step C (1.90 g, 5.46 mmol) and pyridine(948 mg, 12.01 mmol) in CN₂ Cl₂ (30 mL), cooled in an ice bath, wasadded slowly, dropwise thionyl chloride (715 mg, 6.01 mmol). The icebath was removed and the solution stirred at room temperature for 30minutes. Extractive work up and silica gel chromatography (CN₂ Cl₂)afforded 1.35 g (75%) of product as a mixture of olefin isomers. Thematerial was dissolved in EtOAc (20 mL) and 5% Pd on CaCO₃ was added.The mixture was stirred under a balloon of hydrogen for 48 hours. Thecatalyst was removed and the solvent evaporated under reduced pressure.Silica gel chromatography (EtOAc:CN₂ Cl₂ : hexane 3:2:5) afforded 1.35 g(99%) of title product.

¹ N-NMR (300 MHz, CDCl₃) δ1.40-1.85 (m, 7H), 2.12 (m, 1H), 2.54 (dd,J=11 Hz, J=18 Hz, 1H), 2.69 (t, J=7 Hz, 2H), 2.41 (ddd, J=3 Hz, J=7 Hz,J=17 Hz, 1H), 3.17 (m, 1H), 4.40 (m, 1 J), 7.29 (d, J=8 Hz, 2H),7.22-7.40 (m, 5H), 7.57 (d, J=8 Hz, 2H).

E. Preparation of (±)-cis-6- 4-4-(Aminoimino)phenyl)butyl!tetrahydro-4-phenyl-2H-pyran-2-one,trifluoroacetate

The title compound was prepared from the product of step D (1.34 g, 3.51mmol) in a manner similar to example 1, step G affording 720 mg (44%) ofproduct as the TFA salt following reverse phase chromatography (m.p.204°-205° C. dec.).

Anal. calc'd. for C₂₄ H₂₇ N₂ O₄ F₃ 0.25H₂ O: C, 61.46; H, 5.91; N, 5.97.Found: C, 61.33; H, 5.80; N, 6.02.

EXAMPLE 9

Preparation of rel-(4R, 6R)-6-4-(4-(Aminoiminomethyl)phenyl)butyl!-4-hydroxy-4-methyl-3,4,5,6-tetrahydro-2H-pyran-2-one##STR21## A. Preparation of8-(4-cyanophenyl)-4-hydroxy-2-methyl-1-octene.

To a solution of the product of example 1, step C (2.80 g, 15 mmol), andmethylallyl chloride (2.72 g, 30 mmol) in DMF (20 mL) was added NaI(4.50 g, 30 mmol) and zinc dust. The mixture was stirred at ambienttemperature for 2 hours, diluted with aq. 10% HCl and extracted withEtOAc (2×). The organic layer was washed with brine, dried (MgSO₄),filtered and evaporated under reduced pressure. Silica gelchromatography (CH₂ Cl₂) afforded 3.60 g (99%) of title product.

¹ H-NMR (300 MHz, CDCl₃) δ1.35-1.70 (m, 6H), 1.76 (s, 3H), 2.08 (dd, J=9Hz, J=14 Hz, 1H), 2.19 (dd, J=4 Hz, J=14 Hz, 1H), 2.69 (t, J=7 Hz, 2H),3.71 (m, 1H), 4.80 (s, 1H), 4.89 (s, 1H), 7.27 (d, J=8 Hz, 2H), 7.57 (d,J=8 Hz, 2H).

B. Preparation of 8-(4-cyanophenyl)-4-(2-bromo-1-oxoethoxy)-octane-2-one

Ozone was passed through a solution of the product of step A (1.14 g,4.69 mmol) in MeOH (20 mL) at -78° C. until a blue color persisted.After 10 minutes, oxygen was bubbled through the solution until the bluecolor dissipated then dimethylsulfide (1.0 mL) was added and thereaction mixture allowed to warm to room temperature while stirringovernight. The solvent was removed under reduced pressure. To a solutionof the crude residue and pyridine (741 mg, 9.38 mmol) in Et₂ O at 0° C.was added dropwise bromoacetyl bromide (1.04 g, 5.16 mmol). The reactionwas allowed to warm to 15° C. over 1.5 hours then diluted with EtOAc andwashed successively with 1N NaSO₄ and sat'd NaHco₃, dried (MgSO₄),filtered and evaporated under reduced pressure to give 1.00 g of crudeproduct used directly in the next reaction.

¹ H-NMR (300 MHz, CDCl₃) δ1.30-1.75 (m, 6H), 2.17 (s, 3H), 2.55-2.72 (m,3H), 2.79 (dd, J=8 Hz, J=17 Hz, 1H), 3.77 (s, 2H), 5.29 (m, 1H), 7.26(d, J=8 Hz, 2H, 7.57 (d, J=8 Hz, 2H).

C. Preparation of rel-(4R, 6R)-6-4-(4-cyanophenyl)butyl!-4-hydroxy-4-methyl-3,4,5,6-tetrahydro-2H-pyran-2-one

The title compound was prepared from the product of step B (2.66 g, 7.33mmol) in a manner similar to example 8, step C to give 1.88 g (90%) ofproduct after silica gel chromatography (EtOAc:CN₂ Cl₂ l:4).

¹ H-NMR (300 MHz, CDCl₃) δ1.33 (s, 3H), 1.40-1.75 (m, 7H), 1.86 (m, 1H),2.42 (d, J=18 Hz, 1H), 2.60-2.72 (m, 3H), 4.65 (m, 1H), 7.27 (d, J=8 Hz,2H), 7.57 (d, J=8 Hz, 2H).

D. Preparation of rel-(4R, 6R)-6-4-(4-(aminoiminomethyl)phenyl)butyl!-4-hydroxy-4-methyl-3,4,5,6-tetrahydro-2H-pyran-2-one

The title compound was prepared from the product of step C (1.83 g, 6.44mmol) in a manner similar to example 1, step G to give 1 71 g (63%) ofthe title product as the TFA salt following reverse phase chromatography(m.p. 195°-196° C.).

Anal. calc'd. for C₁₉ H₂₅ N₂ O₅ F₃ : C, 54.54; H, 6.02; N, 6.70. Found:C, 54.57; H, 5.99; N, 6.69.

EXAMPLE 10

Preparation of (±)-cis-6- 4-4-(Aminoiminomethyl)phenyl)butyl!tetrahydro-4-methyl-2H-pyran-2-one,trifluoroacetate ##STR22## A. Preparation of 6-4-(4-(aminoiminomethyl)phenyl)butyl!-4-methyl-tetrahydro-2H-pyran-2-one

To a solution of the product of example 9, step D (500 mg, 1.20 mmol) in10 mL of TFA:1,2-dichloroethane (6:4) was added trifluoroaceticanhydride (1.00 g, 4.78 mmol). The solution was stirred at 60° C. for 30minutes, concentrated under reduced pressure. The residue was purifiedby reverse phase chromatography on a Waters® OR C-18 Delta Pak columnusing a 0.05% TFA/water:acetonitrile gradient affording 360 mg (75%) ofproduct (m.p. 188°-191° C. dec.).

Anal. calc'd. for C₁₉ H₂₃ N₂ O₄ F₃ : C, 56.99; H, 5.79; N, 7.00. Found:C, 56.61; H, 5.72; N, 6.95.

B. Preparation of (±)-cis-6- 4-4-(Aminoiminomethyl)phenyl)butyl!tetrahydro-4-methyl-2-H-pyran-2-one,trifluoroacetate

A solution of the product of step A (1.00 g, 2.5 mmol) and 10% Pd/C (50mg) in MeOH (10 mL) was stirred under a balloon of hydrogen for 3 hours.The catalyst was removed and the solvent evaporated under reducedpressure. Recrystallization of the product from acetonitrile afforded560 mg (56%) of product (m.p. 206°-207.5° C. dec.).

Anal. calc'd. for C₁₉ H₂₅ N₂ O₄ F₃ : C, 56.71; H, 6.26; N, 6.96. Found:C, 56.28; H, 6.25; N, 6.95.

EXAMPLE 11

Preparation of (±)-cis-6- 4-4-(Aminoiminomethyl)phenyl)butyl!tetrahydro-4-(3-pyridinyl)-2H-pyran-2-one,ditrifluoroacetate ##STR23## A. Preparation of7-(4-cyanophenyl)-3-hydroxy-1-(3-pyridyl)heptan-1-one

To a solution of diisopropylamine (1.01 g, 10 mmol) in dry THF (20 mL)at 0° C. under an atmosphere of nitrogen was added a 1.6M solution ofnBuLi (6.25 mL, 10 mmol) in hexane. After 15 minutes at 0° C. thesolution was cooled to -78° C. and 3-acetylpyridine (1.21 g, 10 mmol)was added dropwise via syringe. After 10 minutes a solution of theproduct of example 1 step C (1.87 g, 10 mmol) in 10 mL of THF was addedvia canula. The solution was stirred at -78° C. for 30 minutes thenpoured into water (50 mL) and 1N NaHSO₄ (20 mL), extracted with EtOAc,dried (MgSO₄), filtered and evaporated under reduced pressure leaving anoily residue which solidified on standing. Trituration of the residuewith diisopropyl ether gave 1.62 g (53%) of the title product (m.p.93°-97° C.).

Anal. calc'd. for C₁₉ N₂₀ N₂ O₂ : C, 74.00; H, 6.54; N, 9.08. Found: C,73.90; H, 6.80; N, 9.06.

B. Preparation of6-(4-(4-cyanophenyl)butyl-4-(3-pyridyl)-5,6-dihydro-2H-pyran-2-one

To a solution trimethylsilyl P,P-dimethylphosphonoacetate (2.03 g, 8.44mmol) and DMF (5 uL) in CH₂ Cl₂ (10 mL) was added oxalyl chloride (2.45g, 19.47 mmol). The solution was stirred at room temperature until gasevolution ceased (ca. 1 hour). The solvent was removed under reducedpressure and the residue reconstituted with 1,2-dichloroethane andconcentrated again under reduced pressure. This residue of the crudeP,P-dimethyl phosphonoacetyl chloride in THF (10 mL) was added to astirred solution of the product of step A (2.00 g, 6.49 mmol) andpyridine (1.03 g, 13.00 mmol) in THF (60 mL) at 0° C. After 10 minutes,the reaction was removed from the ice bath and stirred at roomtemperature for 1 hour. The reaction mixture was diluted with EtOAc,washed with water, dried (MgSO₄), filtered and concentrated underreduced pressure. The unstable phosphonoacetate ester was dissolved indry THF (50 mL) and cooled to 0° C. under an atmosphere of nitrogen.Solid NaH (60% dispersion in mineral oil) (285 mg, 7.14 mmol) was addedall at once under a stream of nitrogen. The reaction was allowed to warmto room temperature over 1 hour then stirred for an additional 1 hour.The mixture was diluted with EtOAc, washed with 1/2 sat'd NaCl, dried(MgSO₄), filtered and evaporated under reduced pressure. Silica gelchromatography (EtOAc) afforded 1.24 g (58%) of product (m.p. 110°-112°C.).

Anal. calc'd. for C₂₁ H₂₀ N₂ O₂ : C, 75.88; H, 6.07; N, 8.43. Found: C,75.78; H, 6.04; N, 8.31.

C. Preparation ofcis-6-(4-(4-cyanophenyl)butyl-4-(3-pyridyl)-3,4,5,6-tetrahydro-2H-pyran-2-one

To a stirred solution of the product of step B (1.50. g, 4.52 mmol) inEtOAc (30 mL) was added 5% Pd/C (1.50 g) and the mixture stirred under aballoon of hydrogen at room temperature for 20 hours. The reaction wasincomplete as determined by thin layer chromatography, thus anadditional 500 mg of catalyst was added and the reaction stirred for anadditional 20 hours. The catalyst was removed and the solvent evaporatedunder reduced pressure. Silica gel chromatography (EtOAc) afforded 870mg (58%) of the title product.

¹ H-NMR (300 MHz, CDCl₃) δ1.30-1.85 (m, 7H), 2.15 (m, 1H), 2.53 (dd,J=12 Hz, J=18 Hz, 1H), 2.70 (t, J=7 Hz, 2H), 2.94 (ddd, J=2 Hz, J=6 Hz,J=18 Hz, 1H), 3.24 (m, 1H), 4.43 (m, 1H), 7.25-7.35 (m, 3H), 7.53 (dt,J=2 Hz, J=8 Hz, 1H), 7.58 (d, J=8 Hz, 2H), 8.50 (d, J=2 Hz, 1H), 8.55(dd, J=2 Hz, J=5 Hz, 1H).

D. Preparation of (±)-cis-6- 4-4-(Aminoiminomethyl)phenyl)butyl!tetrahydro-4-(3-pyridinyl)-2H-pyran-2-one,ditrifluoroacetate

A solution of the product of step C (100 mg, 0.30 mmol) in dry THF (1mL) was added to a stirred solution of 1.0M lithiumbis(trimethylsilyl)amide (1.2 mL, 1.2 mmol) in dry THF (2 mL) at -78° C.under an atmosphere of nitrogen. The cooling bath was removed and thesolution stirred at ambient temperature for 17 hours. The reaction wascooled in an ice bath and a solution of aq. 6N HCl (0.40 mL, 2.4 mmol)was added. The ice bath was removed and the mixture was stirred atambient temperature for 10 minutes. Evaporation of the solvent underreduced pressure and reverse phase chromatography on a Waters® Delta PakC-18 column using a 0.05% aq. TFA/acetonitrile gradient afforded 60 mg(35%) of product as the bis(TFA) salt m.p. 106° C. (phase change),135°-138° C.!.

Anal. calc'd. for C₂₅ H₂₇ N₃ O₆ F₆ : C, 51.81; H, 4.70; N, 7.25. Found:C, 51.54; H, 4.77; N, 7.26.

EXAMPLE 12

Preparation of N-4-(aminoiminomethyl)phenyl!tetrahydro-6-oxo-2H-pyran-2-propanamide,trifluoroacetate ##STR24## A. Preparation of6-(3-hydroxy-3-oxopropyl)-3,4,5,6-tetrahydro-2H-pyran-2-one

To a mixture of known (cf. A. Ijima et al., 1971)6-(3-butenyl)-3,4,5,6-tetrahydro-2H-pyran-2-one (1.20 0 g, 11.9 mmol)and NaIO₄ (11.50 g, 53.5 mmol) in 88 mL of H₂ O:CCl₄ :CH₃ CN (3:2:2)cooled to -20° C. was added RuCl₃ H₂ O (50 mg, 0.24 mmol). The mixturewas stirred at -20° C. for 2 hours, diluted with water and extractedwith CH₂ Cl₂. The aqueous layer was saturated with NaCl and extractedwith EtOAc (4×). The organic layers were combined, dried (MgSO₄) and thesolvent evaporated under reduced pressure. The residue was filteredthrough a bed of silica gel (EtOAc:Et₂, 1:1) to afford 1.20 g (84%) ofthe title product.

¹ H-NMR (300 MHz, CDCl₃) δ1.50-2.02 (m, 6H), 2.35-2.54 (m, 2H),2.54-2.65 (m, 2H), 4.37 (m, 1H).

B. Preparation of 6-3-(4-cyanophenylamino)-3-oxopropyl!-3,4,5,6-tetrahydro-2H-pyran-2-one

To a solution of the product of step A (900 mg, 7.50 mmol) in CH₂ Cl₂(20 mL) was added oxalyl chloride (4.76 g, 37.5 mmol) and the reactionstirred at room temperature until gas evolution ceased (ca. 1.5 hours).The solvent was removed under reduced pressure and the residueevaporated 2 more times for CH₂ Cl₂ . A solution of the resulting acidchloride in CH₂ Cl₂ (10 mL) was added to a stirred solution of4-aminobenzonitrile (885 mg, 7.50 mmol), dimethylaminopyridine (915 mg,7.50 mmol) and triethylamine (757 mg, 7.50 mmol) in CH₂ Cl₂ (15 mL). Themixture was stirred at room temperature for 18 hours, concentrated underreduced pressure and partitioned between EtOAc and sat'd NaHCO₃. Theorganic layer was washed with 1N NaHSO₄, dried (MgSO₄), filtered andevaporated under reduced pressure. Silica gel chromatography (EtOAc:CH₂Cl₂, 1:1 ) afforded 780 mg (47%) of product (m.p. 145°-148° C.).

Anal. calc'd. for C₁₅ H₁₆ N₂ O₃ :C, 66.16; H, 5.92; N, 10.29. Found: C,65.92; H, 6.02; N, 10.05.

C. Preparation of 6-3-(4-(aminoiminomethyl)phenylamino)-3-oxopropyl!-3,4,5,6-tetrahydro-2H-pyran-2-one

The title compound was prepared from the product of step B (770 mg, 2.83mmol) in a manner similar to example 1, step G to give 430 mg (38%) ofproduct as TFA salt after reverse phase chromatography and triturationwith CH₃ CN/Et₂ O (m.p. 151°-153° C. dec.).

Anal. calc'd. for C₁₇ H₂₀ N₃ O₅ F₃ 0.25N₂ O: C, 50.06; H, 5.07; N,10.30. Found: C, 50.05; H, 4.83; N, 10.45.

EXAMPLE 13

Preparation of 6- 4-(4-(aminoiminomethyl)phenyl)butyl)tetrahydro-2H-pyran-2-one. ##STR25## A. Preparation of 6-4-(4-cyanophenyl)butyl!-3,4,5,6-tetrahydro-2H-pyran-2-one.

A solution of the product of example 4, step A (1.27 g, 0.5 mmol) and 5%Pd on CaCO₃ in EtOAc (50 mL) was stirred under a balloon of hydrogen for2 days. Evaporation and silica gel chromatography (EtOAc:hexane 1:1)afforded 850 mg (67%) of product.

¹ H-NMR (300 MHz, CDCl₃) δ1.3-2.0 (m, 10H), 2.44 (m, 1H), 2.56 (m, 2H),2.68 (t, J=7 Hz, 2H), 4.26 (m, 1H), 7.27 (d, J=8 Hz, 2H), 7.57 (d, J=8Hz, 2H).

B. Preparation of 6-4-(4-(aminoiminomethyl)phenyl)butyl!-3,4,5,6-tetrahydro-2H-pyran-2-one.

The title compound was prepared from the product of step A (850 mg, 3.31mmol) in a manner similar to example 1, step G, affording 710 mg (55%)of product as the TFA salt after reverse phase chromatography andprecipitation from acetonitrile/ether (m.p. 207°-208.5° C.).

Anal. calc'd. for C₁₈ H₂₃ N₂ O₄ F₃ ; C, 55.66; H, 5.97; N, 7.21. Found:C, 55.31; H, 6.12; N, 7.17.

EXAMPLE 14

Preparation of (±)-trans-6-4-(4-(aminiminomethyl)phenyl)-3-butynyl)!-4(4-ethoxyphenyl)-tetrahydro-2H-pyran-2-one.##STR26## A. Preparation of 5-(4-cyanophenyl)-4-pentynal.

The title compound was prepared from the product of example 1, step A(4.39 g, 23.73 mmol) in a manner similar to example 1, step C,substituting 5-(4-cyanophenyl)-4-pentynol for 5-(cyanophenyl)pentanol.Extractive workup afforded 4.40 g (100%) of crude product usedimmediately in the next reaction.

¹ H-NMR (300 MHz, CDCl₃) δ2.70-2.86 (m, 4H), 7.45 (d, J=8 Hz, 2H), 7.57(d, J=8 Hz, 2H), 9.85 (s, 1H).

B. Preparation of (cis/trans)-9-(4-cyanophenyl)3,5-dihydroxy-8-nonynoicacid.

The title compound was prepared from the product of step A (4.34 g,23.73 mmol) in a manner similar to example 1, step D. The residue wasdissolved 50 mL of THF:MeOH (1:1) and 25 mL of 1N NaOH was added. Thereaction mixture was stirred at room temperature for 2 hours, dilutedwith water and extracted with ether. The aqueous phase was acidifiedwith 1N NaHSO₄, extracted with EtOAc, dried (MgSO₄), filtered andevaporated under reduced pressure affording 4.83 g of crude product useddirectly in the next reaction.

¹ H-NMR (300 MHz, CDCl₃) δ1.60-1.85 (m, 4H), 2.50-2.64 (m, 4H),4.07-4.15 (m, 1H), 4.30-4.47 (m, 1H), 7.45 (d, J=8 Hz, 2H), 7.57 (d, J=8Hz, 2H).

C. Preparation of 6-4-(4-cyanophenyl)-3-butynyl!-5,6-dihydro-2H-pyran-2-one.

To a solution of the product of step B (4.60 g, 16.03 mmol) andtriethylamine (9.70 g, 96.16 mmol) in CH₂ Cl₂ (50 mL) at 0° C. was addedmethanesulfonyl chloride (4.57 g, 40.07 mmol). The reaction mixture waswarmed to room temperature and stirred for 2 hours, diluted with CH₂ Cl₂and washed with 1N NaHSO₄, dried (MgSO₄) filtered and evaporated underreduced pressure. Silica gel chromatography (EtOAc/hexane 1:1) of theresidue afforded 850 mg (21%) of product.

¹ H-NMR (300 MHz, CDCl₃) δ1.90-2.02 (m, 1H), 2.04-2.17 (m, 1H), 2.42 (m,2H), 2.70 (m, 2H), 4.62 (m, 1H), 6.07 (dt, J=10 Hz, J=2 Hz, 1H), 6.92(dt, J=10 Hz, J=5 Hz, 1H), 7.45 (d, J=8 Hz, 2H), 7.57 (d, J=8 Hz, 2H).

D. Preparation of (±)-trans-6-4-(4-cyanophenyl)-3-butynyl!-4-(4-ethoxyphenyl)-tetrahydro-2H-pyran-2-one.

To a stirred solution of 4-bromophenetole (2.24 g, 11.15 mmol) in dryTHF (12 mL) at -70° C. under an atmosphere of nitrogen was addeddropwise a 1.6M solution of n-BuLi (6.97 mL, 11.15 mmol) in hexane.After 15 minutes at -70° C., solid CuCN (499 mg, 5.57 mmol) was addedall at once under a stream of nitrogen. The reaction was stirred at -60°C. for 30 minutes then cooled to -70° C. To the resulting solution wasadded the product of step C (700 mg, 2.79 mmol) in THF (5 mL) andstirred at -70° C. for 30 minutes. The reaction mixture was poured intoa rapidly stirred solution of aqueous NH₄ OH/NH₄ Cl, extracted withEtOAc, washed with water, dried (MgSO₄), filtered and evaporated underreduced pressure. Silica gel chromatography (35% EtOAc/hexane) afforded480 mg (46%) of product.

¹ H-NMR (300 MHz, CDCl₃) δ1.42 (t, J=7 Hz, 3H), 1.80-1.91 (m, 1H),2.00-2.13 (m, 3H), 2.65 (t, J=7 Hz, 2H), 2.70-2.87 (m, 2H), 3.35 (pent,J=7 Hz, 1H), 4.00 (q, J=7 Hz, 2H), 4.56 (m, 1H), 6.86 (d, J=8 Hz, 2H),7.12 (d, J=8 Hz, 2H), 7.37 (d, J=8 Hz, 2H), 7.57 (d, J=8 Hz, 2H).

E. Preparation of (±)-trans-6-4-(4-(aminiminomethyl)phenyl)-3-butynyl)!-4-(4-ethoxyphenyl)-tetrahydro-2H-pyran-2-one.

The title compound was prepared from the product of step D (480 mg, 1.29mmol) in a manner similar to example 1, step G affording 283 mg (43%) ofproduct as the TFA salt after reverse phase chromatography andprecipitation from CH₂ Cl₂ /Et₂ O.

Anal. calc'd. for C₂₆ H₂₇ N₂ O₅ F₃ : C, 59.97; H, 5.58; N, 5.38. Found:C, 60.33; H, 5.43; N, 5.29.

EXAMPLE 15

Preparation of (±)-cis-6-4-(4-(aminoiminomethyl)phenyl)butyl!-4-cyclohexyl-3,4,5,6-tetrahydro-2H-pyran-2-one,trifluoroacetate.

A. Preparation of(±)-7-(4-cyanophenyl)-3-hydroxy-1-cyclohexylheptane-1-one.

The title compound is prepared in a manner similar to example 8, step A,substituting cyclohexyl methyl ketone for acetophenone.

B. Preparation of (±)-cis-6-4-(4-aminoiminomethyl)phenyl)butyl!-4-cyclohexyl-3,4,5,6-tetrahydro-2H-pyran-2-one,trifuoroacetate.

The title compound is prepared from the product of step A bysequentially applying the methods described in the following examples:example 8, steps B and C; example 1, step G; example 10, steps A and B.

EXAMPLE 16

Preparation of (±)-cis-6- 4-(4-aminoiminomethyl)phenyl)butyl!-4-(methylsulfonyl)methyl!-3,4,5,6-tetrahydro-2H-pyran-2-one,trifluoroacetate.

A. Preparation of (±)-trans-6- 4-(4-cyanophenyl)butyl!-4-(methylsulfonyl)methyl!-3,4,5,6-tetrahydro-2H-pyran-2-one

The title compound is prepared in a manner similar to example 5, step A,substituting methyl methylsulfonylacetate for methylphenylsulfonylacetate.

B. Preparation of (±)-cis-6- 4-(4-aminoiminomethyl)phenyl)butyl!-4-(methylsulfonyl)methyl!-3,4,5,6-tetrahydro-2H-pyran-2-one,trifluoroacetate.

The title compound is prepared from the product of step A bysequentially applying the methods described in the following examples:example 6, step A; example 4, step C; example 1, step G.

EXAMPLE 17

Preparation of N-4-(aminoiminomethyl)phenyl!3,4,5,6-tetrahydro-6-oxo-4S-hydroxy-2H-pyran-2R-propanamide,trifluoroacetate. ##STR27## A. Preparation of t-butyl5R-hydroxy-9-methyl-3-oxo-8-decenoate.

To a 1M solution of lithium bis(trimethylsilyl)-amide (25.3 mL, 25.3mmol) in THF, cooled to -45° C. was added t-butyl acetate (3.4 mL, 25.3mmol), neat, dropwise via syringe. After 10 minutes, known (c.f. D. F.Taber, L. J. Silverberg, 1991) methyl 3R-hydroxy-7-methyloctanoate (1.47g, 7.90 mmol) in 5 mL of THF was added and stirred at -45° C. for 1hour. The reaction was poured into 1N NaHSO₄ and extracted (2×) withEtOAc, dried (MgSO₄), filtered and evaporated under reduced pressureaffording 2.10 g of crude product used directly in the next reaction.

¹ H-NMR (300 MHz, CDCl₃) δ1.4 (s, 9H) 1.62 (s, 3H), 1.69 (s, 3H), 2.10(q, J=7 Hz, 2H), 2.58-2.77 (m, 2H), 3.39 (2, 2H), 4.07 (m, 1H), 5.11 (t,J=7 Hz, 1H).

B. Preparation of t-butyl 3S,5R-dihydroxy-9-methyl-8-decenoate.

To a solution of tetramethylammonium triacetoxyborohydride (11.83 g, 45mmol) in 60 mL of HOAc/CH₃ CN (1:1), cooled to -30° C. was added theproduct of step A (2.70 g, 10 mmol) in 5 mL CH₃ CN. The reaction wasstirred at -30° C. --20° C. for 2 hours. The reaction was quenced at-30° C. by the addition of 50 mL of 1/2 saturated aqueous Na/K tartrate.After 15 minutes, the stirred solution was covered with 100 mL of Et₂O/hexane (1:1) and NaOH (22 g in 150 mL of H₂ O ) was added slowly. Thelayers were separated and the aqueous layer was extracted with Et₂O/hexane (1:1). The organic layers were combined, washed with saturatedNaCl, dried (MgSO₄), and evaporated under reduced pressure.Crystalization from cold pet. ether afforded 1.50 g of product as awhite solid (m.p. 64.5°-65.5° C.)

Anal. calc'd. for C₁₅ H₂₈ O₄ : C, 66.14; H, 10.36. Found: C, 66.49; H,10.76.

C. Preparation of3S-hydroxy-6R-(4-methylpent-3-enyl)-3,4,5,6-tetrahydro-2H-pyran-2-one.

To a solution of the product of step B (12.55 g, 46.1 mmol) in 20 mL ofMeOH was added NaOH (3.70 g, 92.5 mmol) in 10 mL of water. Afterstirring at ambient temperature for 2 hours, the reaction was pouredinto 100 mL of 1N NaHSO₄. The aqueous phase was saturated with NaCl andextracted (3×) with EtOAc, dried (MgSO₄), and evaporated under reducedpressure. The resulting crude acid was dissolved in benzene and refluxedwith azeotropic removal of water for 4 hours. The solvent was removedunder reduced pressure and the residue chromatographed on silica gel(50-70% EtOAC/hexane) to give 9.10 g (99%) of product.

¹ H-NMR (300 MHz, CDCl₃) δ1.61 (s, 3H), 1.69 (s, 3H), 1.72-1.87 (m, 1H),2.15 (m, 2H), 2.26 (m, 1H), 2.47 (dd, J=16 Hz, J=8 Hz, 1H), 2.90 (dd,J=16 Hz, J=6Hz, 1H), 4.10-4.31 (m, 2H), 5.08 (t, J=7 Hz, 1H).

D. Preparation of3S-(t-butyldimethylsiloxy)-6R-(4-methylpent-3-enyl)-3,4,5,6-tetrahydro-2H-pyran-2-one.

To a solution of the product of step C (9.20 g, 46.5 mmol) in 8 mL ofDMF was added t-butyldimethylsilyl chloride (7.34 g, 48.8 mmol) andimidazole (6.64 g, 97.6 mmol). The reaction was stirred at ambienttemperature for 5 hours, diluted with water and extracted (2×) withhexane. The organic phase was washed with saturated NaCl, dried (MgSO₄),filtered, and evaporated under reduced pressure affording 14.54 g (100%)of crude product used directly in the next reaction.

¹ H-NMR (300 MHz, CDCl₃) δ0.07 (s, 6H), 0.88 (s, 9H), 1.61 (s, 3H), 1.69(s, 3H), 1.72-1.87 (m, 1H), 2.05-2.20 (m, 3H), 2.42 (dd, J=16 Hz, J=8Hz, 1H), 2.81 (dd, J=16 Hz, J=6 Hz, 1H), 4.08-4.22 (m, 2H), 5.08 (t, J=7Hz, 1H).

E. Preparation of3S-(t-butyldimethylsiloxy)-6R-(2-carboxyethyl)-3,4,5,6-tetrahydro-2H-pyran-2-one.

To a stirred mixture of the product of step D (1.00 g, 3.20 mmol) andsodium metaperiodate (3.08 g, 14.40 mmol) in 28 mL of CCl₄ /CH₃ CN/H₂ O(2:2:3) was added 15 mg of RuCl₃.H₂ O. The reaction mixture was stirredvigorously at ambient temperature for 5 hours, diluted with 50 mL ofwater and extracted (2×) with Et₂ O/EtOAc (1:1). The organic phase waswashed with saturated NaCl, dried (MgSO₄), and filtered through a bed ofsilica gel using Et₂ O/EtOAc (1:1) as eluent. Evaporation of the solventunder reduced pressure afforded 940 mg (97%) of crude product as a darkoil used directly in the next reaction.

¹ H-NMR (300 MHz, CDCl₃) δ0.07 (s, 6H), 0.88 (s, 9H), 1.56-1.67 (m, 1H),1.88-2.06 (m, 2H), 2.44 (dd, J=16 Hz, J=8 Hz, 1H), 2.61 (t, J=7 Hz, 2H),2.81 (dd, J=16 Hz, J=6 Hz, 1H), 4.10-4.34 (m, 2H).

F. Preparation ofN-(4-(aminiminomethyl)phenyl!3,4,5,6-tetrahydro-6-oxo-4S-hydroxy-2H-pyran-2R-propanamide,trifluoroacetate.

To a solution of the product of step E (0.50 g, 1.65 mmol), pyridine(140 mg, 1.75 mmol) and 20 mg of 4-dimethylaminopyridine in 3 mL of CH₂Cl₂ was added trimethylsilyl chloride (190 mg, 1.75 mmol). Afterstirring for 15 minutes at ambient temperature, 5 μL of DMF and oxalylchloride (216 mg, 1.70 mmol) were sequentially added. After stirring foran additional 30 minutes at ambient temperature, the reaction mixturewas pipeted into a solution of aminobenzamidine dihydrochloride (343 mg,1.65 mmol) in 3 mL DMF/pyridine (1:1). The reaction mixture was stirredat ambient temperature for 30 minutes, concentrated under reducedpressure and triturated with Et₂ O. After decantation of the Et₂ O fromthe oily residue, the crude material was dissolved in 10 mL of 48% aq.HF/CH₃ CN (1:9) and stirred at ambient temperature for 1 hour. Thesolvent was partially removed under reduced pressure and then trituratedwith Et₂ O. Decantation of the Et₂ O and purification of the residueusing reverse phase chromatography on a Waters® C-18 Delta Pak columnusing a 0.05% TFA/water:acetonitrile gradient afforded 230 mg (33%) ofthe title compound as the TFA salt after trituration with CH₃ CN/Et₂ Om.p. 222.5°-223.5° C. (dec.)!.

Anal. calc'd. for C₁₇ H₂₀ N₃ O₆ F₃.1/3H₂ O: C, 48.01; H, 4.90; N, 9.88.Found: C, 47.82; H, 4.70; N, 9.88.

¹ H-NMR (300 MHz, d₆ -DMSO) δ1.35-1.47 (m, 1H), 1.80-2.02 (m, 2H),2.13-2.31 (m, 2H), 2.40-2.64 (m, 2H), 2.77 (dd, J=16 Hz, J=6 Hz, 1H) ,4.10 (m, 1H) , 4.29 (m, 1H), 7.80 (s, 4H).

EXAMPLE 18

The platelet receptor binding affinity and aggregation inhibitorypotency of representative compounds of the present invention can bedemonstrated by the assays presented below.

In-Vitro Platelet Aggregation in PRP

Healthy male or female dogs were fasted for 8 hours prior to drawingblood; then 30 ml whole blood was collected using a butterfly needle and30 cc plastic syringe with 3 ml of 0.129M buffered sodium citrate(3.8%). The syringe was rotated carefully as blood was drawn to mix thecitrate. Platelet-rich plasma (PRP) was prepared by centrifugation at975× g for 3.17 minutes at room temperature, allowing the centrifuge tocoast to a stop without braking. The PRP was removed from the blood witha plastic pipette and placed in a plastic capped, 50 ml Corning conicalsterile centrifuge tube which was held at room temperature. Plateletpoor plasma (PPP) was prepared by centrifuging the remaining blood at2000× g for 15 minutes at room temperature allowing the centrifuge tocoast to a stop without braking. The PRP was adjusted with PPP to acount of 2-3×10⁸ platelets per ml. 400 μl of the PRP preparation and 50μl of the compound to be tested as prepared below or saline werepreincubated for 1 minute at 37° C. in a BioData aggregometer (BioData,Horsham, Pa.). 50 μl of adenosine 5'diphosphate (ADP) (50 μm finalconcentration) was added to the cuvettes and the aggregation wasmonitored for 1 minute. The test compounds were prepared by thefollowing procedure. The compound was dissolved in 100% ethanol at aconcentration of 10⁻² M. It was vortexed for 30 seconds followed by theaddition of 1.1 equivalents of 1N NaOH. This solution was mixed for 30minutes at room temperature. After the 30 minutes of mixing, thecompound solution was brought to a working concentration of 10⁻³ M withdouble distilled water. All compounds are tested in duplicate. Resultsare calculated as follows:

Percent of control= (maximal OD minus initial OD of compound) divided by(maximal OD minus initial OD of control saline)!×100. The %inhibition=100-(percent of control).

The compounds tested and their median inhibitory concentrations (IC₅₀)are recorded in Table A. IC₅₀ 's (if a compound showed 50% inhibition)were calculated by linear regression of the dose response curve. Theassay results for the representative compounds of the present inventionare set forth in Table A. Also in Table A, two readings given in asingle box indicate that two trials, rather than a single trial, wererun for that particular compound in that particular assay.

                                      TABLE A    __________________________________________________________________________    IN-VITRO PLATELET AGGREGATION    IN PRP                          Dog PRP                          IC.sub.50                                %    Test    Compound              Micro M                                Inhibition                                     Concentration    __________________________________________________________________________    (±)-cis-6- 4- 4-(aminoiminomethyl)                          4.2   100  1 × 10.sup.-5    phenyl!butyl!tetrahydro-4-phenyl-2H-pyran-    2-one, trifluoroacetate    (±)-cis-6- 4- 4-(aminoiminomethyl)                          5.3   89   1 × 10.sup.-5    phenyl!butyl!tetrahydro-4-hydroxy-2H-pyran-                          4.1   96   1 × 10.sup.-5    2-one, trifluoroacetate    (±)-trans-6- 4- 4-(aminoiminomethyl)                                36   1 × 10.sup.-5    phenyl!butyl!tetrahydro-4-hydroxy-2H-pyran-    2-one, trifluoroacetate    (±)-6- 4- 4-(aminoiminomethyl)                                29   1 × 10.sup.-5    phenyl!butyl!tetrahydro-2H-pyran-2-one,                                38   1 × 10.sup.-5    trifluoroacetate    (±)-cis-6- 4- 4-(aminoiminomethyl)                          7.1   74   1 × 10.sup.-5    phenyl)butyl!tetrahydro-4-methyl-2H-pyran-    2-one, trifluoroacetate    (±)-cis-6- 4-(4-(aminoiminomethyl)                          2.6   100  1 × 10.sup.-5    phenyl)butyl!-4-ethenyl-tetrahydro-2H-    pyran-2-one, trifluoroacetate    (±)-cis-methyl 2- 4- 4-(aminoiminomethyl)                          4.0   100  1 × 10.sup.-5    phenyl!butyl!tetrahydro-6-oxo-2H-pyran-4-    acetate    (±)-cis-6- 4- 4-(aminoiminomethyl)                          0.16  100  1 × 10.sup.-5    phenyl)butyl!tetrahydro-4-(3-pyridinyl)-    2H-pyran-2-one, ditrifluoroacetate    (±)cis-6- 4- 4-(aminoiminomethyl)                          2.6   100  1 × 10.sup.-5    phenyl!butyl!tetrahydro-4-     (phenylsulfonyl)methyl!-2H-pyran-2-one,    trifluoroacetate    (±)-trans-6- 4- 4-(aminoimino)phenyl)butyl!                                7    1 × 10.sup.-5    tetrahydro-4-phenyl-2H-pyran-2-one,    trifluoroacetate    N- 4-(aminoiminomethyl)phenyl!tetrahydro-6-                          2.9    oxo-2H-pyran-2-propanamide,    trifluoroacetate    N- 4-(aminoiminomethyl)Phenyl!tetrahydro-6-                          0.35    oxo-4S-hydroxy-2H-pyran-2-propanamide,    trifluoroacetate    __________________________________________________________________________

What we claim is:
 1. A compound of the formula ##STR28## or apharmaceutically acceptable salt thereof, wherein R₃ and R₄ are eachindependently selected from the group consisting of hydrido, alkylhaving 1 to 6 carbon atoms, hydroxy, alkoxy having 1 to 6 carbon atomsand halo;X is --CH₂ CH₂ --, --CH═CH--, --C.tbd.C-- or HNCO m is aninteger from 1 to 3; and A is the group ##STR29## wherein R₁ is hydrido;hydroxy; alkyl having 1 to 6 carbon atoms; alkenyl having 2 to 6 carbonatoms which may be optionally substituted by halo; alkynyl having 2 to 6carbon atoms; alkoxycarbonyalkyl; phenylsulfonyalkyl;alkylsulfonylalkyl; phenyl which may be optionally substituted by alkylhaving 1 to 6 carbon atoms, alkoxy having 1 to 6 carbon atoms andhydroxy; or cycloalkyl having 3 to 6 carbon atoms.
 2. A compoundaccording to claim 1 wherein R₁ is hydrido.
 3. A compound according toclaim 1 wherein X is --CH₂ CH₂ -- or --CH═CH--.
 4. A compound accordingto claim 1 which is (±)-cis-6- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4-phenyl-2H-pyran-2-one,trifluoroacetate.
 5. A compound according to claim 1 which is (±)-cis-6-4- 4-aminoiminomethyl)phenyl!butyl!tetrahydro-4-hydroxy-2H-pyran-2-one,trifluoroacetate.
 6. A compound according to claim 1 which is(±)-trans-6- 4- 4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4-hydroxy-2H-pyran-2-one, trifluoroacetate.
 7. A compound according to claim 1 whichis (±)-6- 4- 4-(aminoiminomethyl)phenyl!butyl!tetrahydro-2H-pyran-2-one,trifluoroacetate.
 8. A compound according to claim 1 which is N-4-(aminoiminomethyl)phenyl!tetrahydro-6-oxo-2H-pyran-2-propanamide,trifluoroacetate.
 9. A compound according to claim 1 which is (±)-cis-6-4- 4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4-methyl-2H-pyran-2-one,trifluoroacetate.
 10. A compound according to claim 1 which is(±)-cis-6- 4-4-(aminoiminomethyl)phenyl!butyl!-4-ethenyltetrahydro-2H-pyran-2-one,trifluoroacetate.
 11. A compound according to claim 1 which is(±)-cis-methyl 2- 4- 4-(aminoiminomethyl)phenyl!butyl!tetrahydro-6-oxo-2H-pyran-4-acetate.
 12. A compoundaccording to claim 1 which is (±)-cis-6- 4- 4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4- (phenylsulfonyl) methyl!-2H-pyran-2-one,trifluoroacetate.
 13. A compound according to claim 1 which is(±)-trans-6- 4-4-(aminiminomethyl)phenyl!butyl!tetrahydro-4-phenyl-2H-pyran-2-one,trifluoroacetate.
 14. A compound according to claim 1 which is N-4-(aminoiminomethyl)phenyl!3,4,5,6-tetrahydro-6-oxo-4S-hydroxy-2H-pyran-2R-propanamide,trifluoroacetate.
 15. A pharmaceutical composition useful for inhibitingplatelet aggregation comprising an effective amount of a compoundaccording to claim 1 together with one or more non-toxicpharmaceutically acceptable carriers.
 16. A pharmaceutical compositionaccording to claim 15 wherein the compound is selected from the groupconsisting of(±)-cis-6- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4-phenyl-2H-pyran-2-one,trifluoroacetate; (±)-cis-6- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4-hydroxy-2H-pyran-2-one,trifluoroacetate; (±)-trans-6- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4-hydroxy-2H-pyran-2 -one,trifluoroacetate; (±)-6- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-2H-pyran-2-one,trifluoroacetate; (±)-cis-6- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4-methyl-2H-pyran-2-one,trifluoroacetate; (±)-cis-6- 4-4-(aminoiminomethyl)phenyl!butyl!-4-ethenyltetrahydro-2H-pyran-2-one,trifluoroacetate; (±)-cis-methyl 2- 4- 4-(aminoiminomethyl)phenyl!butyl!tetrahydro-6-oxo-2H-pyran-4-acetate; (±)-cis-6- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4-(phenylsulfonyl)methyl!-2H-pyran-2-one, trifluoroacetate; (±)-trans-6-4- 4-(aminiminomethyl)phenyl!butyl!tetrahydro-4-phenyl-2H-pyran-2-one,trifluoroacetate; N-4-(aminoiminomethyl)phenyl!tetrahydro-6-oxo-2H-pyran-2-propanamide,trifluoroacetate; and N-4-(aminoiminomethyl)phenyl!3,4,5,6-tetrahydro-6-oxo-4S-hydroxy-2H-pyran-2R-propanamide,trifluoroacetate.
 17. A method of treating a mammal to inhibit plateletaggregation comprising administering a therapeutically effective dose ofa compound of claim 1 to a mammal in need of said treatment.
 18. Amethod according to claim 17 wherein said compound is selected from thegroup consisting of(±)-cis-6- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4-phenyl-2H-pyran-2-one,trifluoroacetate; (±)-cis-6- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4-hydroxy-2H-pyran-2-one,trifluoroacetate; (±)-trans-6- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4-hydroxy-2H-pyran-2-one,trifluoroacetate; (±)-6- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-2H-pyran-2-one,trifluoroacetate; (±)-cis-6- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4-methyl-2H-pyran-2-one,trifluoroacetate; (±)-cis-6- 4-4-(aminoiminomethyl)phenyl!butyl!-4-ethenyltetrahydro-2H-pyran-2-one,trifluoroacetate; (±)-cis-methyl 2- 4- 4-(aminoiminomethyl)phenyl!butyl!tetrahydro-6-oxo-2H-pyran-4-acetate; (±)-cis-6- 4-4-(aminoiminomethyl)phenyl!butyl!tetrahydro-4-(phenylsulfonyl)methyl!-2H-pyran-2-one, trifluoroacetate; (±)-trans-6-4- 4-(aminiminomethyl)phenyl!butyl!tetrahydro-4-phenyl-2H-pyran-2 -one,trifluoroacetate; N-4-(aminoiminomethyl)phenyl!tetrahydro-6-oxo-2H-pyran-2-propanamide,trifluoroacetate; N-4-(aminoiminomethyl)phenyl!3,4,5,6-tetrahydro-6-oxo-4S-hydroxy-2H-pyran-2R-propanamide,trifluoroacetate.